Manual Biology 2016 (9642016B)

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MAJLIS PEPERIKSAAN MALAYSIA

(MALAYSIAN EXAMINATIONS COUNCIL)
PEPERIKSAAN
SIJIL TINGGI PERSEKOLAHAN MALAYSIA
(MALAYSIA HIGHER SCHOOL CERTIFICATE EXAMINATION)
Manual for
Biology Coursework
Paper 4 (964/4)
STPM 2016
REMINDER:
This manual is specifically for the use of teachers or examiners only and should not be given to
unauthorised persons.
________________________________________________________________________________________________________________
This manual consists of 125 printed pages.

© Majlis Peperiksaan Malaysia 2015
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Contents
Page
Part 1: Teacher’s Manual for Biology Coursework
1.1 Introduction 1
1.2 General Information 1
1.3 Recording of Assessment Marks 2
1.4 Moderation 3
1.5 Practical Work Assessment Guide 4
1.6 Title of the experiment 6
1.7 Table of Summary of Experiment 7
1.8 Marking scheme/Teacher guide 8
1.9 List of Apparatus and Materials 32
Part 2: Student’s Manual for Biology Coursework
2.1 Introduction 45
2.2 Assessment of Practical Work 46
2.3 Title of the experiment 48
2.4 Experiment for First Term 49
2.5 Experiment for Second Term 75
2.6 Experiment for Third Term 102
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BIOLOGY COURSEWORK STPM 2016
Part 1: Teacher's Manual for Biology Coursework
1.1 Introduction
1.1.1 This manual contains administration and guidelines on the implementation and
assessment of the practical work which have to be carried out by the school.
1.1.2 Some of the skills that should be developed in STPM subject (e.g. handling of
apparatus, observation, interpretation of results, and planning) can only be fully
acquired through practical work.
1.1.3 Continuous assessment of practical work in school throughout form six will ensure that
direct assessment of all the desired practical and scientific skills of students can be
made.
1.1.4 The practical science assessment is carried out in schools with the following objectives.
(a) To establish a practical work assessment system which is fair, accurate and
comprehensive
(b) To improve the practical skills and the quality of practical work of students
(c) To inculcate independence, teamwork spirit, scientific attitudes and critical
thinking among students
1.2 General Information
1.2.1 The teacher in charge of the school-based assessment of practical biology will be
provided with a softcopy of the Teacher’s Manual which contains the details of the
administration of practical biology assessment, practical work assessment guide and
description of experiments.
1.2.2 Malaysian Examinations Council (MEC) will provide a softcopy of Teacher's and
Student’s Manual which can be downloaded from MEC Portal
(http://www.mpm.edu.my) during the first term of form six. MEC will sent username
and password to Principal of the school for the downloaded purposes. The school is
expected to make the duplicate copies of the Student's Manual which contain
experiments to be given to each student.
1.2.3 MEC has determined 15 compulsory experiments to be carried out by students which
will be assessed by the teacher in three respective terms.
1.2.4 Experiments are to be carried out either individually or in groups as recommended in

the Table of Summary of Experiments on pages 7.
1.2.5 The teacher is expected to prepare the experiments according to this manual. MEC
should be informed of any modifications made by using the Experiment Report Form
(See Appendix B on page 44).
1.2.6 The recommended time to complete the practical works report is as recommended in
the Table of Summary of Experiments on pages 6.
1.2.7 The information on each experiment should be given to the students before the
experiment is carried out so that they can plan their practical work.
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1.2.8 The teacher should ensure that a student has been given a chance to acquire a particular
skill before the assessment of that skill is made. For this purpose, the teacher should
carry out similar experiments before carrying out the compulsory experiments.
1.2.9 The assessment of experimental skills should be done while the student is carrying out
the experiment and also on the student’s practical work report.
1.2.10 For a student who is absent for an experiment with reason, the teacher can fix another
date for the students to carry out the experiment.
1.2.11 Students may write their practical work reports in either English or Bahasa Melayu.
The practical work reports is to be submitted to the teacher on the same day the
experiment is carried out unless otherwise stated. (Refer to the Table of Summary of
Experiments on pages 7). Practical work reports which are not submitted on the day of
the experiment are to be awarded ‘0’ mark.
1.2.12 Practical work reports which can be completed at home are to be submitted to the
teacher not later than 3 days from the date of the experiment. A penalty of 2 marks is to
be imposed for the reports submitted late to the teacher. Practical work reports which
are submitted later than 7 days from the date of the experiment are to be awarded ‘0’
mark.
1.2.13 For a student who has transferred to another school, the previous school is to send the
student’s reports and the Student Record which is partially completed and signed by the
subject teacher to the student’s new school.
1.2.14 All practical work and Student's Record are evidence; and should be kept by the school
and destroyed under secure condition 6 months after the release of the STPM result in
the following year.
1.2.15 All practical work are part of the STPM Biology 964 syllabus and can be assess as
part of written exam.
1.3 Recording of assessment marks
1.3.1 Recording of the practical assessment marks of each student is to be done by the subject
teacher on Student's Record (See Appendix A on page 43).
1.3.2 For each student, the teacher is to record the date of the experiment, the experiment
number and mark given to each of the skills for the 15 compulsory experiments in the
Student's Record. Marks are to be awarded in accordance with the Marking Scheme/
Teacher’s Guide for experiments on pages 8 to 31.
1.3.3 Remarks on the students for the following cases should be written in the ‘Notes’
column on the Student's Record.
(a) An experiment carried out at a later date for a student who was absent for the
experiment
(b) Any penalty imposed due to late submission of practical reports to the teacher
(c) A student who has not finished all the experiments allocated (reasons to be stated)
(d) A student who failed to submit all the practical (reasons to be stated)
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1.3.4 Practical work for the three terms should be completed four weeks before the written
examination for third term.
1.3.5 Once the practical assessment for every term is completed, the marks for the
15 compulsory experiments is to be calculated and written in the ‘Overall Total mark’
column on the Student's Record. The full total mark for this practical work is 225.
1.3.6 The total mark for each student must be submitted to MEC via electronic submission in
the specified date.
1.3.7 The teacher carrying out the practical assessment are required to make a declaration that
the entries of marks and the overall total mark in the Student's Record are correct by
signing in the spaces provided.
1.4 Moderation
1.4.1 A common assessment standard for marking must be agreed upon if more than one
teacher in the same school is involved in assessing the student's practical work to ensure
that the internal assessment is carried out fairly and effectively.
1.4.2 All component of assessment are subjected to moderation by the moderator appointed
by MEC.
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1.5 Practical Work Assessment Guide
Assessment
Theme Description Marks
criterion
Microscope Manipulative • Skill in handling microscope

and Slide skill
• Ability to follow instructions
2−4
• Skill in preparation and displaying
specimens
Result • Skills in drawing/accuracy, labels,

5–8
scale, and identification specimens
Discussion • Ability to answer questions correctly 2–6
Conclusion • Ability to make relevance conclusion 1–2
Biochemistry Manipulative • Preparation of materials, procedures,

and Physiology skill including preparation of solution and
experimental substances
• Planning and execution (structuring, 3–6
planning and managing, neatness and
following instructions)
• Skill in observation and drawing
Result • Ability to label and plot a graph

correctly and accurately
• Ability to perform calculation correctly 3–6
and accurately
• Record reading with precision
Dissection Manipulative • Ability to follow instructions on how to

skill dissect and display a system
• Accuracy and completeness of display 3–4
• Neatness of dissection
• Ability to identify the organ
Result • Skill in drawing and labels the organ

5–6
and system of the specimen
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Assessment
criterion
Ecology Manipulative • Skill in collection and preservation of

skill specimens
• Achieving target (number and accuracy
of identification)
2–8
• Quality of specimen and preservation
• Skill in handling equipment
Result • Skill in labeling and displaying

specimens
and accurately
• Diversity (family and order)
Genetic Manipulative • Ability to follow instruction

skill 2–4
Result • Perform correct calculations

4–5
• Skill in drawing and labeling
Discussion • Ability to answer questions correctly 6
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1.6 Titles of Experiments
Topic in Time
Experiment Working
the Title of Experiment (2 hours =
number Mode
syllabus 3 periods)
First Term
2 1 Measurement of cell size using 2 hours Individual
microscope
2 2 Preparation and observation of 2 hours Individual
slides of animal and plant cells
3 3 Determination of the osmotic 2 hours Individual
potential of plant cells
4 4 Effect of temperature on enzymatic 2 hours Individual
activity
5 5 Oxygen consumption during 2 hours Group
respiration in plant
6 6 Separation of photosynthetic 2 hours Individual
pigments using paper
chromatography
6 7 Comparison between C3 and C4 2 hours Individual
leaves
Second Term
7 8 Dissection of the mammalian 2 hours Individual
respiratory system
8 9 Dissection of the mammalian blood 2 hours Individual
circulatory system
10 & 11 10 Dissection of the mammalian urino- 2 hours Individual
genital system
10 11 Embryogenesis in animal 2 hours Individual
11 12 Examination of slides: kidney, liver 2 hours Individual
and pancreas
Third Term
14 13 Classification of organisms using a 2 hours Individual
dichotomous key
14 14 Preservation techniques 2 hours Individual
14 15 Collections of insects of 10 3weeks Group
different orders and plants of 10
different families
15 16 Ecological study of a terrestrial 3weeks Group
habitat
17 17 Mendelian inheritance 2 hours Individual
17 18 Extraction of plant DNA 2 hours Individual
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BIOLOGY COURSEWORK
STPM 2016
1.7 Table of Summary of Experiments
Group of Biology Microscope and slide Biochemistry and Dissection Ecology Genetic
Practical titles physiology
A Compulsory 1; 2; 7; 11; 12 3; 4; 5; 6 8; 9; 10 13; 14; 15; 16 17; 18

practicals [5 experiments] [4 experiments] [3 experiments] [4 experiments] [2 experiments]
[18 experiments]
B Compulsory 1; 2; 11; 12 4; 5; 6 8; 9; 10 13; 15; 16 17; 18

practicals [4 experiments] [3 experiments] [3 experiments] [3 experiments] [2 experiments]
[15 experiments]
Notes:
A Compulsory practicals which must be carried out.
B Compulsory practicals to be used for assessment purposes.
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1.8 Marking Scheme/Teacher’s Guide for Experiments
1.8.1 Experiment 1
Manipulative skills (A) Marks
− Focusing 1
− Slide preparation (neat + no air bubble) 1+1
3
Results (B)
Part A
1. Drawing and labelling of Amoeba/Paramecium (3 Label) 1
2. Eye piece & objective lens magnification 1
3. Apparent object size & size of drawing 1
Part B
1. Diameter microscope field of view 2
2. No of cell lengthways 1
3. No of cell widthways 1
7
Discussion (C)
Part A
1. Magnification of Amoeba/Paramecium drawing 1
2. Actual Amoeba/Paramecium size 1
Part B
1. Average length 1
1. Average width 1
4
Conclusion (Suggested) (D)

Different cells vary in sizes 1
1
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TERHAD
1.8.2 Experiment 2
− Preparation of blood smear 1
− Preparation of thin stem section 1
− Focusing clearly from low magnification to medium magnification 1
3
Results (B)
Drawing, labelling and magnification
− Blood cells (RBC, WBC) 1
− Plan diagram of dicot stem (any 3 labels) 1+1
− Detailed diagram of dicot stem (any 3 labels) 1+1
5
Discussion (C)
1. (a) Circular/sphere in shape 1
(b) RBC no nucleus, WBC with nucleus 1
WBC is larger than RBC//RBC spherical WBC irregular shape 1
2. Methylene blue stained the nucleus of the WBC 1
3. Minimum 3 types of cells 1
4. Metaxylem absent//only protoxylem present//individual vascular bundle
arrange in a ring at the side of the stem 1
6
Conclusion (suggested) (D)

Animal cells are different from plant cells 1
1
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TERHAD
1.8.3 Experiment 3
− Correct equipment used 1
− Correct way of using equipment 1
− Follow instruction 1
3
Results (B)
(a) Dilution of solution
Table 2: Volume of the stock solution and distilled water used in each dilution.
Sucrose concentrations (M) 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

Volume of 1.0 M sucrose solution
2 4 6 8 10
used (ml)
Volume of distilled water used (ml) 18 16 14 12 10
− Correct volume of stock solution and distilled water used as shown

in the table above. 1
(b) Tabulation result

Table 3:
Test tube 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

Average initial length of 3 strips (mm)
Average final length of 3 strips (mm)
Average change in length of strips
(mm)
Average initial weight of 3 strips (g)
Average final weight of 3 strips (g)
Average change in weight of 3 strips (g)
Texture of strips at the end of
experiment
− Average length of strips (fill up at least 3 columns) 1
− Average weight (fill up at least 3 columns) 1
− Texture of strips (fill up at least 3 columns) 1
(c) Plotting of graphs

(i) A plotted graph shows the average change in the length of the
potato strips against the concentrations of the sucrose solution
used – (a slight curve) 1
(ii) A plotted standard graph shows the osmotic potential against the
concentrations of the sucrose solution – (a straight line graph) 1
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The graphs – correct title, axes, and the plot
Change in length
(mm)
0 Concentrations
0.2 0.4 0.6 0.8
(M)
Discussion (C)
(a) (i) The concentration where the curve crosses the x-axis in Graph 2 1
Explanation:
When the average change in length of the potato strips is zero,
there is no net movement of water in and out of the potato cells//
The solute concentration / osmotic potential of the potato cell sap
is isotonic to the surrounding sucrose solution. 1
(ii) The osmotic potential 1
(b) Increase in weight the texture becomes harder (turgid)//

decrease in weight the texture becomes softer (flaccid). 1
(c) Over dose of fertilizer will create a hypertonic condition/will decrease osmotic
potential outside the plant cells//Water will be drawn out of the
cell by osmosis and cause the plant to wilt and die. 1

From the experiment, the osmotic potential of potato cell sap
is determined (the value). 1
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1.8.4 Experiment 4
Manipulative skill (A) Marks
− Preparation of saliva solution and set up apparatus 1
(test-tubes, water baths at the right temperatures)
− The way the experiment is carried out and tidiness 1
− The way observation is carried out 1
Results (B)
− Results tabulated 1
− Graph correctly plotted:
• x−axis and y−axis correct, labels of axes with unit are correct
• Correct plotting of point
• Smooth and correct curve 2/0
(all three must be correct) 3
Discussion (C)
(1) Temperature at the peak of the graph 1
(2) − Enzyme denatures/destroy the tertiary structure of enzymes
− By breaking bonds (hydrogen bonds, ionic bonds) between polypeptide chain
− Alter shape of active site
− Substrates unable to bind with enzymes
− Fewer enzyme-complexes formed
− Rate of reaction decreases/stop
(any 5) 5
(3) (i) Temperature coefficient = ~2 (accept 1 − 3) 1

(ii) A value of 2 for Q10 means that the rate of reaction doubles for
each 10 ºC rise in temperature 1
8

As the temperature increases, the rate of enzyme reaction increases until
it reaches the optimum temperature, it then decreases and stop at 60 ºC 1
1
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1.8.5 Experiment 5
− Preparation of tubes 1
− Adding drop of graduated pipette 1
− Marker 1
3
Results (B)
− Data tabulation (Initial position, table, total volume) 3
− Graph correctly plotted:
• x-axis and y-axis correct, labels of axes with unit are correct
• Correct plotting of point
• Smooth and correct curve (all three must be correct) 2/0
5
Discussion (C)
1. Oxygen is used/consumed 1
2. (a) To absorb the CO2 1
(b) Change in volume maybe small/insignificant 1
− Carbon dioxide released compensate oxygen consumed 1
3. (x/820) × (1g)
− Correct answer 1
4. Atmospheric pressure and temperature 1
6
Conclusion (D)
The amount of oxygen consumption is.... 1
1
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1.8.6 Experiment 6
Manipulative skill (A) Marks
− Correct way of using mortar and pestle 1
− Chromatography paper − 1
pointed end
width
neatness
pigment spot preparation
− Apparatus set-up − 1
position of large test tube
position of strip
solvent level
− Cleanliness of table and safety 1
(all solvent containers are tightly stopped)
4
Results (B)
Green Blue− Yellow Yellow Orange

Pigments colour
green − grey
Distance moved by pigment / mm
Distance moved by solvent / mm
Rf value 0.45 0.65 0.71 0.83 0.95
(1 mark each for distance and Rf value) 1+1

Chromatogram with marked and label 1
3
Discussion (C)
1. Chlorophyll b = green 1
Chlorophyll a = blue-green 1
Xanthophyll = yellow 1
Phaeophytin = grey
Carotene = orange max 3
2. Size, solubility and the adsorption (any 2) 1
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3. (a) (answer depends on the student’s result) 1

− Smallest in size
− Highest in solubility
− Least adsorption (any 2) 1
(b) The smaller the Rf value the larger the size//
the higher the Rf value the smaller the size 1
7

Chlorophyll consists of five different pigments that are chlorophyll b,
chlorophyll a, xanthophyll, phaeophytin and carotene. 1
1
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1.8.7 Experiment 7
Focusing − Specimen can be seen clearly (1 mark for each C3 and C4) 1
Slide − intact 1
2
Results (B)
Plan drawing
− Drawing and labelling for C3 leaf + magnification 1+½
High power drawing

6
Discussion (C)
1. C3 – mesophyll cell 1
C4 – bundle sheath cell 1
2. C3 – mesophyll cell 1
C4 – mesophyll cell 1
3. Place where Calvin cycle occurs/rich in Rubisco//chloroplast less grana 1
4. C4 concentrates CO2 in the bundle sheath cells
(where most of Rubisco present)//reduced photorespiration 1
6
Conclussion (suggested) (D)

– C3 and C4 have different leaf (anatomy) structures//
C4 plant shows Krantz anatomy 1
1
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1.8.8 Experiment 8

− Ability to follow instructions on how to dissect and display respiratory system.
(i) Ability to follow instructions 2
(ii) Ability to identify the organ accurately 1
− Tidiness/neatness 1
4
Results (B)
− Drawing and labelling (larynx, heart, trachea and lungs) 1+4
5
Disscussion (C)
1. − upward and outward movements of the rib cage increase the volume of
thoracic cavity 1
− Downward and inward movements of the rib cage decrease the volume
of thoracic cavity 1
2. (a) Thin and flexible 1
(b) Ability to change shape in accordance to change the volume
of thoracic cavity 1
3. Nose/mouth → larynx → trachea → bronchus → bronchiole → alveolus 1
5

− Rat internal respiratory system consists of larynx, lungs, trachea,
diaphragm and rib cage 1
1
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1.8.9 Experiment 9
− Ability to follow instructions on how to dissect and display blood circulatory
system.
(ii) Ability to identify the organs accurately 1
4
Results (B)
− Drawing 1 + label
(Pulmonary vein, pulmonary artery, aorta,
right atrium, right ventricle − any 3) 1+1
(innominate artery, right subclavian artery,
right carotid artery − all 3) 1+1
(right atrium, right ventricle, left atrium, left ventricle − all 4) 1+1
6
Discussion (C)
1. To supply (oxygenated) blood to neck and head/upper part of the body 1
2. Low oxygen content//deoxygenated blood//wider diameter//thinner wall 1
3. Deoxygenated blood enter right atrium → right ventricle. 1
Oxygenated blood enter left atrium → left ventricle 1
4
Heart is divided into 4 chambers. 1
Rat blood circulatory system consists of heart, veins and arteries.
1
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1.8.10 Experiment 10
− Ability to follow instructions on how to dissect and urino-genital system
(ii) Ability to identify the organs accurately 1
3
Results (B)
− Male + label (any 3 urino + any 3 genital) 1+2
− Female + label (any 3 urino + any 3 genital) 1+2
6
Discussion (C)
1. Synthesis/release of aldosterone hormone 1
controls osmolarity of blood (plasma) 1
2. Accessory organs 1
3. Male: to release urine and sperm 1
Female: to release urine only 1
5

− Male and female rats have different urino-genital system 1
1
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Results (B) (drawing) Marks
− 4-celled 1
− Morula 1
− Blastocyst and 3 label 1+2
− Gastrula and 3 label 1+2
8
Discussion (C)
1. Sperm and ovum 1
2. Gives rise/Develop into chorion/placenta 1
3. To form cell layers//precursor for cell layers 1
4. (i) Bone, cartilage, muscle, adipose tissue, connective tissue,
blood vascular, reproductive, excretory and urinogenital systems,
notochord, mesothelium, spleen, heart, blood vessel, dermis 1
(ii) Gastrointestinal tract (alimentary canal, pharynx,
terminal part of rectum) Respiratory tract (trachea, bronchi, alveoli)
Endocrine glands and organ (lining of thyroid gland and thymus)
Auditory system (epithelium of auditory tube and tympanic cavity)
Urinary system (bladder, part of urethra) 1
(iii) Nervous system (brain, spine, peripheral nerve), tooth (enamel),
epidermis, Lining of mouth, anus, nostril, sweat gland, hair, nail 1
6

− Embryogenesis begins with a single cell called zygote/ 1
Cell differentiation starts after blastula/
Three germ layers give rise to different organs (or tissues) 1
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Instruction to teacher:
Show the diagram of embryo at 4-celled, morula, blastocyst and gastrula stages given below to the
students for them to draw and label.
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− Focusing and correct ways of using equipments 1
− Follow instructions 1
2
Results (B)
1. Plan drawing of kidney with 3 label (cortex, medulla, pelvis) 1
2. Renal cortex drawing + simple squamous and cuboidal epithelial 1+1
3. Lobule structure drawing 2
4. Portal area drawing 2
5. Islet of Langerhans drawing with 2 label 1
8
Discussion (C)
1. Tubules are made of simple cuboidal epithelial cells 1
2. In blood vessels//hepatic artery/vein//sinusoid 1
3. Artery has thicker wall/small lumen//vein has thinner wall/
larger lumen 1
4. Many tubules in the kidney (less compact) 1
4

− Kidney, liver and pancreas have different cellular organizations//
kidney has many tubules, liver has many lobules
and pancreas has islet of cells. 1
1
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− Use of equipment 1
− Observation skill 1
2
Results (B)
− Spider key 4
− Dichotomous key 4
8
Discussion (C)
1. (a) Segmented body with exoskeleton 1
Appendage/jointed legs 1
(b) Arthropoda 1
2. Marchantia Presence of gemmae cups//Funaria spore found in the capsule

Dryopteris Presence of sori 1
4

− Dichotomous keys are used for classify organism 1
1
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Dichotomus Key
A1 Organism containing chlorophyll........................................................................... refer to B
A2 Organism without chlorophyll………………………….................……............... refer to D
B1 Organism does not produce spores.

Organism has roots, stems and leaves. grass
.......……..................................................................................................................
B2 Organism produces spores..........................…….................................................... refer to C
C1 Organism has leaves (fronds)

sub-divided into pinnae and pinnules
bearing sori on undersides……………………..................……………………… Dryopteris
C2 Organism with thalloid body,
has gemma cups, antheridiophores
and archegoniophores as
reproductive structures................……………….................................................. Marchantia
//Leaf like structure, seta and capsule………………………………………… Funaria
D1 Unicellular organism......................……………………………………………..... Amoeba
D2 Multicellular organism............................................................................................ refer to E

E1 Organism without exoskeleton but has soft cylindrical body.
Body wall contains two layers of cells
− ectoderm and endoderm
Has tentacles and a gut cavity with a single
opening called a mouth……….....................…………………………………….. Hydra
E2 Organism with exoskeleton…………………………………............................…. refer to F
F1 Invertebrate, has soft muscular foot,

and calcareous shell...........................................................………………………. Snail
F2 Invertebrate with jointed legs...............................................................…………... refer to G
G1 Invertebrate, body divided into

head, thorax and abdomen.
Has three pairs of legs, wingless...............................................................……….. Ant
G2 Invertebrate, body divided into

cephalothorax and abdomen.
Has two pairs of antennae
and five pairs of legs............................…………………………………..………. Prawn
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Spider key
Organisms
A1 A2
Organism with chlorophyll Organism without chlorophyll
(Marchantia, Dryopteris, Grass) (Amoeba, Hydra, Ant, Snail, Prawn)
B1 B1 D1 D2
Organism does not Organism produce spores Unicellular organism Multicellular organism
produce spores (Grass) (Marchantia, Dryopteris) (Amoeba) (Hydra, Ant, Snail, Prawn)
C1 C2 E1 E2
Organism has leaves Organism does not have Organism without Organism with exoskeleton
(Dryopteris) leaves (Marchantia) exoskeleton (Hydra) (Ant, Snail, Prawn)
F1 F2
Organism with jointed Organism without jointed
leg (Ant, Prawn) leg (Snail)
G1 G2
Body divided into head, thorax Body divided into
and abdomen cephalothorax and abdomen
(Ant) (Prawn)
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1. Good quality specimens
− Dry house fly 1
− Wet house fly 1
− Dry dragonfly 1
− Wet Marchantia / Funaria 1
− Dry Dryopteris 1
2. Follow instructions 1
6
Results (B)
Labelled specimens
− Dry house fly 1
− Wet house fly 1
− Dry dragonfly 1
− Wet Marchantia / Funaria 1
− Dry Dryopteris 1
5
Discussion (C)
1. As preservative 1
2. Drying or soaking may cause the color to fade 1
3. Closing spiracles may block air passage into the body 1
3
Conclusion (Suggested) (D)

Animal and plant specimens can be preserved for long term//
storage for further studies. 1
1
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− Skill in collecting the samples 3
− Correct preservation method (plant + insect) 1+1
− Quality of preservation (plant + insect) 1+1
− Labelling according to format (plant + insect) 1+1
9
Results (B)
− The way specimens are displayed (plant + insect) 1+1
− 10 orders 2
(if less than 10 orders – 1 mark)
− 10 families 2
(if less than 10 families – 1 mark)
6
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− Correct technique of using quadrat 2
− Naming the plant species 1
− pH determination 1
− Soil collection 1
5
Results (B)
− Table 1 + Calculation 1+1
− Table 4 1
7
Discussion (C)
1. To precipitate clay//to make liquid clear 1
2. Species x is most suitable to grow in pH y (average) 1
2

Plant with the highest species frequency/species density/
species coverage is the most dominant species in the habitat 1
1
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− Follow instruction 1
− Ears of corn remain in good condition 1
2
Results (B)
− Table 2 (2 lines = 1 mark, χ2 =1 marks) 1+1
− Table 3 (2 lines = 1 mark, χ2 =1 marks) 1+1+1
5
Discussion (C)
Part A:
1. 1 1
2. Most numbered phenotype-dominant 1
Least numbered phenotype-recessive 1
3. χ2 0.05 =3.841 1
4
Conclusion (D)
χ2 value less than χ2 0.05, obey Mendel`s First Law 1
1
Discussion (C)
Part B:
1. 3 1
2. χ2 0.05 =7.815 1
2
Conclusion (D)
χ2 value less than χ2 0.05, obey Mendel`s Second Law 1
1
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− Crushing skill (do not produce excessive bubbles) 1
− Ability to obtain / transfer 2 ml of aqueous phase into a new tube 1
− Properly mixing of the mixture with circular motion 1
− Ability to obtain DNA 1
4
Results (B)
Drawing
− Drawing 1
− indicate the presence of DNA at the tips of pooling stick 1
Describing
− physical characteristics of DNA sticky 1
− colourless / whitish in colour 1
4
Discussion (C)
1. Nucleus 1
2. Breaking down of cell wall 1
3. Precipitate DNA 1
4. Electrophoresis 1
5. Nucleotides 1
6. Interphase 1
6
Conclusion (some suggestions) (D)

− Living cells posses DNA//
Breaking the cell wall releases DNA//
DNA in solution can be precipitated and visualized 1
1
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1.9 List of Apparatus and Materials
1.9.1 List of Apparatus and Materials by experiments
Experiment No.
Apparatus/Materials Quantity used
(Mode of working)
Onion bulb 1
Iodine solution C
Light microscope C
1
Glass slides C
(Individual)
Prepared slide of Amoeba 1
Cover slips C
Forceps C
Safranin / light green staining solution 10 cm3
Stem of young dicotyledonous plant (Helianthus
sp. or any soft stem plant)
Blood (to be taken during the class)
Methylene blue 10 cm3
Ethanol (70%)
Ethanol (100%)
Cotton balls 1
2 Light microscope 1
(individual) Slides 1
Cover slips 1
Forceps 1
Mounted needle 1
Petri-dish (9 cm) 1
Fine brush 1
Filter paper strips 1
Sterile disposable lancet 1
Razor blade 1
Note: “C” means common use.
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Experiment No.
(Mode of working)
Sucrose solution 1.0 M 50 cm3
Distilled water C
Potato 1
Large test tubes 5
Beaker 1
Ruler 1
Filter paper 1
Petri dish with cover C
3 Marker pen 1
(individual) Cork-borer 1
Single-edge blade 1
White tile 1
Graph paper 1
Dissecting needle / Mounted needle 1
Measuring cylinder 25 ml 1
Fine forceps 1
Pipette 10 ml 1
Analytical balance 1
Beaker 250 ml 3
Thermometer 10
Test-tube 10
White tile 1
Dropper 2
4 Stopwatch 1
(individual) Starch solution 1% 30 cm3
Iodine solution 30 cm3
Measuring cylinder 10 ml 2
Distilled water C
Hot water C
Ice cube C
Mung bean seeds 200 pieces
Volumeter (readily assembled before class) 1
Glass beads
Non-absorbent cotton 1
5
Potassium hydroxide (KOH) pellets 10g
(Group)
Marker fluid in dropping bottle 2
Labeling sticker 2
Vaseline 1
Retort stand 2
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Experiment No.
(Mode of working)
Acetone 80% 5 cm3
“Cekur manis” or Red spinach leaves 5 pieces
Mortar and pestle 1
Muslin (cheese cloth) 1
Large test tube with stopper (24 × 150 mm) 1
Solvent (1 part 80% acetone and 9 parts petroleum 5 cm3
6 ether)
(individual) Ruler 1
Whatman no. 1 chromatography paper strip 1
(20 × 130 mm)
Scissors 1
Pin 1
Dark paper 1
Retort stand/test tube rack 1
Light microscope C
Prepared slide of transverse section of Zea mays 1
7
leaf
(Individual)
Prepared slide of transverse section of Helianthus 1
sp. leaf
Rat 1
Dissecting instruments 1
Dissecting board 1
8 Thread 1
(Individual) Dissecting pins 15
Diethyl ether 5 cm3
Killing jar 1
Cotton balls 1
Rat 1
Dissecting board 1
Magnifying glass (× 10) 1
9
Thread 1
(Individual)
Dissecting pins 15
Diethyl ether 5 cm3
Killing jar 1
Cotton balls 1
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Experiment No.
(Mode of working)
Rat 1
Dissecting board 1
Magnifying glass (× 10) 1
10
Thread 1
(Individual)
Dissecting pins 15
Diethyl ether 5 cm3
Killing jar 1
Cotton balls 1
CD of embryogenesis 1
11 Computer 1
(Individual) LCD projector 1
Projector screen 1
Prepared microscope slides of kidney 1
Prepared microscope slides of liver 1
12
Prepared microscope slides of pancreas 1
(Individual)
Light microscope C
Lens paper 1
Light microscope C
Magnifying glass 1
Amoeba slide 1
Hydra slide 1
13 Ant 1
Individual) Snail 1
Prawn 1
Marchantia/Funaria 1
Dryopteris 1
Grass 1
Formalin acetic acid (FAA) solution 5 cm3
Ethanol 70% 100 cm3
Diethyl ether 100 cm3
Insect pins 15
Polystyrene board 1
Insect box 1
14
Labeling papers 10
(individual)
Test tubes 50 ml 10
Specimen bottles 5 ml 10
Specimen bottles 50 ml 10
Petri dishes 5
Cotton wool or cotton balls
Newspapers
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Experiment No.
(Mode of working)
Magnifying glass 1
Drying oven 1
Live insects: house fly (small insect) and dragonfly 1
14
(large insect)
(individual)
Plant specimens: fresh fern leaf of Dryopteris and
liverwort Marchantia/Funaria
Naphthalene balls 1
For insect
Insect pooter or aspirator 1
Insect net 1
Collecting tubes 1
Killing jar 1
Magnifying glass 1
4 − 10% formalin
Tray 1
White paper 1
Insect pin 1
15
Insect box 1
(Group)
Naphthalene balls 1
For plant
Plastic bags 1
Thread 1
Newspapers 10 pieces
Plant press 1
Measuring tape 1
Plant cutter 1
Herbarium paper (Drawing block) 1
Needle 10
1 m × 1 m quadrat 10
Test-tubes with stopper 5
Test-tube rack 1
Spatula 1
10 cm3 pipette 1
16 Universal indicator C
(Group) Barium sulphate powder
Soil sample
pH chart C
Soil borer 1
Plastic bags 5
Marker pen 1
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Experiment No.
(Mode of working)
Corns: purple and yellow seeds 1
17 Corns: smooth and wrinkled seeds 1
(individual) Corns: purple / yellow and smooth/wrinkled seeds 1
Pins 10
Ice cold isopropanol 10 ml
Onion (Indian onion gives better result) 1
18 DNA extraction kit (which contains homogenising 1set
(Individual) sticks, extraction buffer, conical centrifugation
tubes, plastic pipette)
Knife 1
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1.9.2 List of apparatus and materials by experiments
Quantity Experiment Mode of

No Apparatus/Material
used no. working
1 Acetone 80% 5 cm3 6 Individual
2 Amoeba slide 1 13 Individual
3 Analytical balance 1 3 Individual
4 Ant 1 13 Individual
5 Barium sulphate powder 16 Group
6 Beaker 250 cm3 3 4 Individual
7 “Cekur manis” or red spinach leaves 5 pieces 6 Individual
8 CD of embryogenesis 1 11 Individual
9 Collecting tube 1 15 Group
10 Computer 1 11 Individual
11 Cork-borer 1 3 Individual
12 Corns: purple and yellow seeds 1 17 Individual
Corns: smooth and wrinkled seeds 1 17 Individual
Corns: purple / yellow and smooth / 1 17 Individual
wrinkled seeds
13 Cotton balls 1 8 Individual
1 9 Individual
1 10 Individual
14 Individual
14 Cotton wool or cotton balls 14 Individual
15 Cover slips C 1 Individual
2 Individual
16 Dark paper 1 6 Individual
17 Diethyl ether 5 cm3 8 Individual
5 cm3 9 Individual
5 cm3 10 Individual
100 cm3 14 Individual
18 Dissecting board 1 8 Individual
1 9 Individual
1 10 Individual
19 Dissecting instruments 1 8 Individual
1 9 Individual
1 10 Individual
20 Dissecting needle / Mounted needle 1 2 Individual
1 3 Individual
21 Dissecting pins 15 8 Individual
15 9 Individual
15 10 Individual
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used no. working
22 Distilled water C 3 Individual
C 4 Individual
23 DNA extraction kit (which contains 1 set 18 Individual
homogenizing sticks, extraction buffer,
conical centrifugation tubes, plastic
pipette)
24 Dropper 2 4 Individual
25 Drying oven 1 14 Individual
26 Dryopteris 1 13 Individual
27 Ethanol 70% 2 Individual
100 cm3 14 Individual
28 Filter paper 1 3 Individual
29 Fine forceps 1 3 Individual
30 Forceps C 1 Individual
C 2 Individual
31 Formalin acetic acid (FAA) solution 5 cm3 14 Individual
32 Glass beads 5 Group
33 Glass slides C 1 Individual
2 Individual
34 Glass tube/Cappilary tube 3mm diameter 2 5 Group
35 Graph paper 1 3 Individual
36 Grass 1 13 Individual
37 Herbarium paper (Drawing block) 1 15 Group
38 Hot water C 4 Individual
39 Hydra slide 1 13 Individual
40 Ice cold isopropanol 10 ml 18 Individual
41 Ice cube C 4 Individual
42 Insect box 1 14 Individual
1 15 Group
43 Insect net 1 15 Group
44 Insect pin 15 14 Individual
1 15 Group
45 Insect pooter or aspirator 1 15 Group
46 Iodine solution C 1 Individual
C 2 Individual
30 cm3 4 Individual
47 Killing jar 1 8 Individual
1 9 Individual
1 10 Individual
1 15 Group
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used no. working
48 Knife 1 18 Individual
49 Labeling papers 10 14 Individual
50 Labeling sticker 2 5 Individual
51 Large test tube with stopper 1 6 Individual
(24 × 150 mm)
52 Large test tubes 5 3 Individual
53 LCD projector 1 11 Individual
54 Lens paper C 2 Individual
1 12 Individual
55 Light microscope C 1 Individual
C 2 Individual
C 7 Individual
C 12 Individual
C 13 Individual
56 Live insects: house fly (small insect) and 1 14 Individual
dragonfly (large insect)
57 Magnifying glass 1 8 Individual
1 9 Individual
1 10 Individual
1 13 Individual
1 14 Individual
1 15 Group
58 Marchantia 1 13 Individual
59 Marker fluid in dropping bottle 2 5 Group
60 Marker pen 1 3 Individual
1 16 Group
61 Measuring cylinder 10 ml 2 4 Individual
62 Measuring cylinder 25 ml 1 3 Individual
63 Measuring tape 1 15 Group
64 Mortar and pestle 1 6 Individual
65 Mung bean seeds 40 pieces 5 Group
66 Muslin (cheese cloth) 1 6 Individual
67 Naphthalene balls 1 14 Individual
1 15 Group
68 Needle 10 15 Group
69 Newspapers C 14 Individual
C 15 Group
70 Non-absorbent cotton 1 5 Group
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used no. working
71 Onion bulb 1 1 Individual
1 18 Individual
72 Petri dish with cover C 3 Individual
C 14 Individual
73 pH chart C 16 Individual
74 Pin 1 6 Individual
10 17 Individual
75 Pipette 10 ml 1 3 Individual
1 16 Individual
76 Plant cutter 1 15 Group
77 Plant press 1 15 Group
78 Plant specimens: fresh fern leaf of 1 14 Individual
Dryopteris and liverwort
Marchantia/Funaria
79 Plastic bags 1 15 Group
80 Polystyrene board 1 14 Individual
81 Potassium hydroxide (KOH) pellets 10 5 Group
82 Potato 1 3 Individual
83 Prepared microscope slides of kidney 1 12 Individual
Prepared microscope slides of liver 1 12 Individual
Prepared microscope slides of pancreas 1 12 Individual
84 Prepared slide of Paramecium, Amoeba or 1 1 Individual
Euglena 1 2 Individual
85 Prepared slide of transverse section of 1 7 Individual
Helianthus sp. leaf
Prepared slide of transverse section of Zea 1 7 Individual
mays leaf
86 Projector screen 1 11 Individual
87 1 m × 1 m quadrat 10 16 Individual
88 Rat (Rabbit optional) 1 8 Individual
1 9 Individual
1 10 Individual
89 Retort stand 2 5 Group
1 6 Individual
90 Ruler 1 3 Individual
1 6 Individual
91 Sand C 5 Group
92 Scissors 1 6 Individual
93 Shrimp 1 13 Individual
94 Single-edge blade 1 3 Individual
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used no. working
95 Snail 1 13 Individual
96 Soil borer 1 16 Group
97 Soil sample 16 Group
98 Solvent (1 part of 80% acetone and 9 5 cm3 6 Individual
parts of petroleum ether)
99 Spatula 1 16 Group
100 Specimen bottles 5 ml 10 14 Individual
101 Specimen bottles 10 ml 10 14 Individual
102 Starch solution 1% 30 cm3 4 Individual
103 Stopper 5 16 Group
104 Stopwatch 1 4 Individual
105 Sucrose solution 1.0 mol 50 cm3 3 Individual
106 Syringe 2 5 Group
107 Test-tube 50 ml 10 4 Individual
10 14 Individual
5 16 Group
108 Test-tube rack 1 16 Group
109 Thermometer 10 4 Individual
110 Thread 1 8 Individual
1 9 Individual
1 10 Individual
1 15 Group
111 Tray 1 15 Group
112 Universal indicator C 16 Group
113 Vaseline 1 5 Group
114 Whatman no. 1 chromatography paper 1 6 Individual
strip (20 × 130 mm)
115 White paper 1 15 Group
116 White tile 1 3 Individual
1 4 Individual
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Appendix A
Student’s Record
BIOLOGY COURSEWORK
PAPER 4 (964/4) STPM 2016
Name of school/institute
Name of student
Centre no./Index no. S / I/C no.
Mark for skill assessed

Skill
A B C D PLD
Marks
Study Teacher’s Teacher’s Moderator’s Teacher’s Moderator’s Teacher’s Moderator’s Level
Date Experiment
term mark mark mark mark mark mark mark
Experiment 1
Experiment 2
First Experiment 4
Experiment 5
Experiment 6
Experiment 8
Experiment 9
Second Experiment 10
Experiment 11
Experiment 12
Experiment 13
Experiment 15
Third Experiment 16
Experiment 17
Experiment 18
Overall Total Mark
TEACHER’S DECLARATION MODERATOR’S DECLARATION

I certify that the marks and details recorded above are true. I certify that the marks and details recorded above are true.
……………………………………………………………… ………………………………………………………………
Name: Name:
Date: Date:
Contact number (H/P): Contact number (H/P):
Contact number (O): Contact number (O):
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Appendix B
BIOLOGY COURSEWORK
PAPER 4 (964/4) STPM 2016
EXPERIMENT REPORT
Name of school/college: …………………………………………………………………………………………………. Centre number: ………………………….
Experiment
Topic Problem/Modification/Suggestion/Comment
Number
Note: If there is no problem/modification/suggestion/comment for a certain experiment, please write “NONE”.

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Part 2: Student’s Manual for Biology Coursework
2.1 Introduction
2.1.1 MEC has determined 15 compulsory experiments to be carried out by students which
will be assessed by the teacher in three respective terms.
2.1.2 Experiments are to be carried out either individually or in groups as recommended in

the Titles of Experiments on pages 48.
2.1.3 The assessment of experimental skills should be done while the student is carrying out
the experiment and also on the student’s practical work report.
2.1.4 For a student who is absent for an experiment with reason, the teacher can fix another
date for the students to carry out the experiment.
2.1.5 Students may write their practical work reports in either English or Bahasa Malaysia.
The practical work reports is to be submitted to the teacher on the same day the
experiment is carried out unless otherwise stated. Practical work reports which are not
submitted on the day of the experiment are to be awarded ‘0’ mark.
2.1.6 Practical work reports which can be completed at home are to be submitted to the
teacher not later than 3 days from the date of the experiment. A penalty of 2 marks is to
be imposed for the reports submitted late to the teacher. Practical work reports which
are submitted later than 7 days from the date of the experiment are to be awarded ‘0’
mark.
2.1.9 For a student who has transferred to another school, the previous school is to send the
student’s reports and the Student Record which is partially completed and signed by the
subject teacher, to the student’s new school.
2.1.10 All practical works are part of the STPM Biology 964 syllabus and can be assess as
part of written exam.
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TERHAD
2.2 Assessment of Practical Work
Assessment
criterion
Microscope Manipulative • Skill in handling microscope

and Slide skill
2−4
• Skills in preparation and displaying
specimens
Result • Skill in drawing/accuracy, labels, scale,

5–8
and identification specimens
Biochemistry Manipulative • Preparation of materials, procedures,

and Physiology skill including preparation of solution and
experimental substances
• Planning and execution (structuring, 3–6
planning and managing, neatness and
following instructions)
• Skill in observation and drawing
Result • Ability to label and plot a graph

correctly and accurately
and accurately
• Record reading with precision
Dissection Manipulative • Ability to follow instructions on how to

skill dissect and display a system
• Accuracy and completeness of display 3–4
• Neatness of dissection
• Ability to identify the organ
Result • Skill in drawing and labels the organ

5–6
and system of the specimen
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Assessment
criterion
Ecology Manipulative • Skill in collection and preservation of

skill specimens
• Achieving target (number and accuracy
of identification)
2–8
• Quality of specimen and preservation
• Skills in handling equipment
Result • Skill in labeling and displaying

specimens
and accurately
• Diversity (family and order)
Genetic Manipulative • Ability to follow instruction

skill 2–4
Result • Perform correct calculations

4–5
• Skill in drawing and labeling
Discussion • Ability to answer questions correctly 6
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2.3 Titles of Experiments
Topic in Time
Experiment Working
the Title of Experiment (2 hours =
number Mode
syllabus 3 periods)
First Term
2 1 Measurement of cell size using 2 hours Individual
microscope
2 2 Preparation and observation of 2 hours Individual
slides of animal and plant cells
3 3 Determination of the osmotic 2 hours Individual
potential of plant cells
4 4 Effect of temperature on enzymatic 2 hours Individual
activity
5 5 Oxygen consumption during 2 hours Group
respiration in plant
6 6 Separation of photosynthetic 2 hours Individual
pigments using paper
chromatography
6 7 Comparison between C3 and C4 2 hours Individual
leaves
Second Term
7 8 Dissection of the mammalian 2 hours Individual
respiratory system
8 9 Dissection of the mammalian blood 2 hours Individual
circulatory system
10 & 11 10 Dissection of the mammalian urino- 2 hours Individual
genital system
10 11 Embryogenesis in animal 2 hours Individual
11 12 Examination of slides: kidney, liver 2 hours Individual
and pancreas
Third Term
14 13 Classification of organisms using a 2 hours Individual
dichotomous key
14 14 Preservation techniques 2 hours Individual
14 15 Collections of insects of 10 3weeks Group
different orders and plants of 10
different families
15 16 Ecological study of a terrestrial 3weeks Group
habitat
17 17 Mendelian inheritance 2 hours Individual
17 18 Extraction of plant DNA 2 hours Individual
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2.4 Experiment for First Term
Experiment 1
Title: Measurement of cell size using microscope
Objective: To measure the size of onion cell and Amoeba/Paramecium using microscope
Learning outcome:
At the end of the experiment, student should be able:
(a) to learn the correct technique of using light microscope;
(b) to estimate the magnification of a drawing made with the help of a microscope;
(c) to estimate the actual size of onion cell and Amoeba/Paramecium;
(d) to determine the size of onion scale leaf epidermal cells.
Introduction
A microscope is a basic tool to observe microscopic specimens or microorganisms. It is a
valuable precision optical instrument if used correctly and carefully. A compound microscope
consists of an illuminator, a condenser, a diaphragm, a stage, two focusing knobs, a nosepiece
and a series of lenses (ocular and objective) which focus radiation passing from the specimen to
form an image. The image usually stands out against its background (contrast) and enlarged
(magnification) but still preserving the specimen details (resolution). Observation of a
specimen should always start with low (4/10×), medium (40/60×) and finally high-power
(100×) magnifications. However, observation of specimen at 100× magnification must be
done under oil immersion. The area of the field of view decreases dramatically with
increasing magnification.
Materials and apparatus

Microscope
Slides of Amoeba/Paramecium
Glass slide
Coverslip
Forceps
Onion
Iodine solution
Part A
To estimate the magnification of a drawing made with the help of a microscope.
1. Examine the slides of various types of microorganisms under a high power microscope.
2. Estimate the image size (apparent object size) of the specimen.
3. Make a labelled drawing and determine the magnification of your drawing using the
following formula:
Magnification Size of drawing
= Magnification of × Magnification of ×
of drawing eyepiece objective lens Apparent size of object
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4. Estimate the actual size of each microorganism using the following formula.
Apparent size of object

Actual size of object =
Magnification of microscope
Results
Drawing
Data
Eye piece magnification = ..................................................………………………………...........

Objective lens magnification = .................................................…………………………………
Microorganism Amoeba/Paramecium
Apparent object size
Size of drawing
Discussion
1. Determine the magnification of the drawing of Amoeba/Paramecium.
2. Estimate the actual size of Amoeba/Paramecium in μm.
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Part B
To determine the size of onion scale leaf epidermal cells.
1. Using a pair of forceps, peel off the epidermal layer of the onion scale leaf.
2. Mount the epidermal layer in a drop of water on a slide.
3. Stain the specimen with iodine solution. Cover the slide with a cover slip.
4. Determine the diameter of the microscope’s field of view by placing a clear plastic ruler
under the microscope and focusing on the millimeter scale at low power. Estimate the
diameter of the microscope`s field of view. Convert it to μm and tabulate your
observations.
Data
To determine the diameter of a microscope’s field of view.
Diameter of low power field of view = ..........................................................................................
Diameter of medium power field of view = ...................................................................................
Diameter of high power field of view = .........................................................................................
Magnification of high power objective lens = ................................................................................
5. Examine the epidermal cells under microscope using high power objective lens.
6. Count the number of cells across the high power field of view length ways. Repeat three
times and determine the average number of cells.
7. Repeat step 6, but this time count the number of cells across the high power field of view
widthways.
8. Calculate the average length and width of a single epidermal cell. State your answer in μm.
Results
Number of cells lengthways:
First count = ................................................... Average = ............................……………

Second count = ...............................................
Third count = ..................................................
Number of cells widthways:

First count = .................................................... Average = ..................................…….........
Second count = ................................................
Third count = ...................................................
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Discussion
1. What is the average length of one epidermal cell of the onion scale leaf?
Average length of one epidermal cell of onion scale leaf

Diameter of field of view used
=
Number of cells lengthways
=
2. What is the average width of one epidermal cell of the onion scale leaf?
Average width of one epidermal cell of onion scale leaf
Diameter of field of view used

=
Number of cells widthways
Conclusion
..................................……...............................................................................................................
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Experiment 2
Title: Preparation and observation of slides of animal and plant cells
Objective: To differentiate between animal and plant cells
Learning outcome:
At the end of the experiment, students should be able:
(a) to prepare slides of blood cells using smear technique;
(b) to prepare slides of plant stem using sectioning technique;
(c) to identify the specialized cells found in blood and plant stem.
Introduction
All organisms are composed of cells, the basic unit of structure and functions. The cell is the
lowest level of structure capable of performing the activities of life. Animal and plant cells are
eukaryotic cells. In general, animal cell is irregular in shape with a central nucleus which may
be lobed. It has compact and granular cytoplasm. On the other hand, a plant cell is more regular
in shape. It has a peripheral nucleus and a compact cytoplasm. A microscope is an important
tool in studying different cell structures and its variations. We can observe clearly the structures
of cells by using an appropriate staining solutions. In this experiment, Safranin/light green
staining solution is used to stain cytoplasm and cellulose (green), nucleus and lignin (red) and
chloroplast (pink).
Material and apparatus

Safranin/light green staining solution
Stem of a young dicotyledonous plant (Helianthus sp. or any soft stem plant)
Blood (to be taken during the practical class)
Methylene blue
Ethanol (70%)
Ethanol (100%)
Cotton balls
Light microscope
Slides
Cover slips
Forceps
Mounting needles
Petri dish (9 cm)
Fine brush
Filter paper strips
Sterile disposable lancet
Razor blade
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Procedure
Part A: Preparation and observation of blood cells
1. Clean the tip of the middle finger with a cotton ball soaked in ethanol (70%).
2. Press the tip of the finger with a thumb and prick it with a sterile disposable lancet.
Caution: The lancet must not be used by more than one person and must be disposed after
use.
3. Squeeze the finger with a thumb to ooze a drop of blood.
4. Place the drop of blood onto one end of a clean, dry slide.
5. Place a second slide on the first slide as shown in Figure 1 and allow the blood to spread
across the region of contact by capillary action.
This slide is pressed down and slide against the

lower one in the direction of the arrow shown
A drop of blood is made to

spread between two slides
Making a blood smear for examination under a

microscope. The blood smear will be stained with
methylene blue to show the nuclei of leucocytes
Figure 1
6. Press and push the top slide forward to allow a thin film of blood smear formed. Remove
the top slide.
7. Let the blood smear to dry at room temperature.
8. Fix the blood smear with 100% ethanol for 1 minute. Drain off excessive ethanol.
9. Stain the blood smear with methylene blue for 5 to 10 minutes.
10. Drain off the excessive methylene blue using a filter paper strip and let it dry at room
temperature. (Note: Do not use cover slip)
11. Examine the slide under low (4/10×), medium (40/60×) objective lens.
12. Identify and draw the different types of blood cells.
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Part B: Preparation and observation of plant stem

1. Pour 10 ml Safranin/light green staining solution into a Petri dish.
2. Hold a dicotyledonous stem between thumb and index finger. Position the thumb a little
higher than index finger.
3. Place a sharp razor blade to rest on your index finger of the same hand that holds the stem.
4. Cut the stem as thin as possible by moving the razor blade towards thumb rapidly.
Caution: Be careful not to cut your thumb with the sharp razor blade.
5. Place all stem sections into a Petri dish containing Safranin/light green stain for
5 minutes.
6. Use a fine brush to dish out the thinnest (the most transparent) stem section and mount it in
a drop of water on a slide.
7. Examine the slide under low power (4/10×) objective lens and draw a plan diagram of the
section.
8. Examine the slide under medium power (40/60×) objective lens and draw a detailed
diagram of a sector in the section.
Magnification Magnification Magnification Size of drawing

= × ×
of drawing of eyepiece of objective lens Apparent size of object
Results
(a) Drawing of blood cells
Drawing magnification: ……………………………….
(b) Drawing of plant stem

(i) Plan diagram
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(ii) Detailed diagram
Discussion
Part A
1. (a) State the appearance of the red blood cell as seen under the microscope.
.........................................................................................................................................................
(b) State two differences between red blood cells and white blood cells as seen under the
microscope.
.........................................................................................................................................................
.........................................................................................................................................................
.........................................................................................................................................................
2. What is the function of the methylene blue solution in this experiment?
.........................................................................................................................................................
Part B
3. Name the types of cells found in the dicotyledonous stem.
.........................................................................................................................................................
4. State the typical characteristic of a young dicotyledonous stem in Helianthus.
.........................................................................................................................................................
Conclusion
.........................................................................................................................................................
.........................................................................................................................................................
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Experiment 3
Title: Determination of the osmotic potential of plant cells
Objective: To determine the osmotic potential of the potato cell sap
Learning Outcome:
At the end of the experiment, student should be able:
(a) to prepare solutions of various concentrations from a stock solution;
(b) to tabulate the results and plot graphs;
(c) to relate between solute concentrations and water potential.
Introduction
Osmotic potential of a cell refers to the amount of solute dissolved in the cell sap, containing
mostly water and contribute to its osmotic concentration. The difference in osmotic
concentration inside the cell and its surrounding is called a concentration gradient. Movement
of water molecules via semi permeable membrane occurs from a region of high water
concentration (low osmotic potential) to a region of low water concentration (high osmotic
potential).

1.0 M sucrose solution
Distilled water
Potato
Large test tubes/ Boiling tubes
Beakers
Ruler
Filter paper
Petri dish with cover
Marker pen
Cork-borer
Single-edge blade
White tile
Graph paper
Dissecting needle / Mounted needle
25 ml measuring cylinder
Fine forceps
10 ml pipette
Analytical balance
Procedure
1. Plot a standard curve (Graph 1) of the osmotic potential against the concentrations of the
sucrose solutions using the data provided in Table 1.
Table 1: Relationship between sucrose concentration and osmotic potential.
Sucrose concentration
0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55
(M)
Osmotic potential
1.3 2.6 4.0 5.3 6.7 8.1 9.6 11.1 12.6 14.3 16.0
(in atmosphere)
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2. In labelled test tubes, prepare 20 ml of sucrose solutions of 0.1 M, 0.2 M, 0.3 M, 0.4 M,
and 0.5 M from the stock solution using the dilution technique. Use the following formula
to assist you to do the dilution.
M1V1 = M2V2 ,where M1 = initial concentration

V1 = initial volume
M2 = final concentration
V2 = final volume
3. Record the volume of the stock solution and of the distilled water used in each dilution in
Table 2.
4. Prepare 15 cylindrical strips of potato tissues using a cork-borer. Cut the strips to a uniform
length.
5. Take 3 potato strips, record their average initial length and weight and place them into the
test tube containing 0.1 M sucrose solution.
6. Repeat step 4 using 0.2 M, 0.3 M, 0.4 M and 0.5 M sucrose solutions.
7. After 30 minutes, remove the strips with the dissecting needle/mounted needle provided.
Wipe them dry gently with the filter paper and record their average final length and weight.
8. Note down any changes to the texture of the potato strips. Tabulate your results in Table 3.
9. Plot another graph (Graph 2) of the average change in the length of the potato strips against
the concentrations of the sucrose solution used.
10. Use both the graphs to determine the osmotic potential of the potato cell sap.
Results
Table 2: Volume of the stock solution and distilled water used in each dilution
Concentrations (M) 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

Volume of 1.0 M sucrose solution used (ml)
Volume of distilled water used (ml)
Table 3: Changes in length, weight and texture of potato strips
Test tube 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

Average initial length of 3 strips (mm)
Average final length of 3 strips (mm)
Average change in length of strips (mm)
Average initial weight of 3 strips (g)
Average final weight of 3 strips (g)
Average change in weight of 3 strips (g)
Texture of strips at the end of experiment
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Discussion
1. (a) From Graph 2, determine the solute concentration of the potato cell sap. Explain your
answer.
.........................................................................................................................................................
.........................................................................................................................................................
.........................................................................................................................................................
(b) From the standard curve (Graph 1), determine the equivalent osmotic potential
(in atmosphere) of the cell sap.
.........................................................................................................................................................
2. What is the relationship between the changes in weight of the potato strips and their
texture?
.........................................................................................................................................................
.........................................................................................................................................................
.........................................................................................................................................................
3. Plant fertilizer contains many different solutes. Why an overdose of fertilizer may kill a
plant?
.........................................................................................................................................................
.........................................................................................................................................................
Conclusion
.........................................................................................................................................................
.........................................................................................................................................................
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Experiment 4
Title: Effect of temperature on enzymatic activity
Objective: To investigate the effects of temperature on the rate of amylase reaction
Learning outcome:
(a) to understand the effect of temperature on the enzyme-catalysed reactions;
(b) to understand the concept of optimal temperature on the enzymatic reactions;
(c) to understand the concept of temperature quotient, Q10, of an enzyme-controlled reaction.
Introduction
Enzymes are complex chemicals that control the reactions in living cells. They are biochemical
catalysts that speed up the rate of metabolic reactions, where larger molecules are broken into
smaller ones or small molecules are built up into more complex ones. Since all enzymes are
globular proteins, the amino acid chain of an enzyme is folded into a globular shape. The
globular shape of enzyme molecule is held in its tertiary form and three dimensional shapes by
hydrogen bonds, ionic bonds and disulphide bridges. Changes in conditions, such as pH and
temperature cause the three dimensional shape of the protein to change. When enzymes react,
they combine with their substrates to form enzyme-substrate complexes. When the reaction has
completed, the products are released, leaving the enzyme in its original form ready to be reused.
Enzymes are fast acting as they have a high turnover number meaning that they can convert
many molecules of substrate per unit time. Up to certain extend heating can increase the rate of
most enzymatic reactions. The effect of temperature on the rate of enzyme reaction can be
expressed by the temperature coefficient Q10:
Rate of reaction at (T + 10) °C
Q10 =
Rate of reaction at T °C
Material and Apparatus

3 units of 250 ml beaker
Thermometer
10 units of test-tube
White tile
Dropper
Stopwatch
30 ml of 1% starch solution
Iodine solution
2 units of 10 ml measuring cylinder
Distilled water
Hot water
Ice cube
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Procedure
Thermometer
A2 A1
Test tube
Ice at 0 ºC
5 ml of starch 1 ml of saliva
suspension
Figure 1: Assembling of enzyme activity tubes
1min 2 3 4 5
6 7 8 9 10
Figure 2: White tile for iodine test
1. Prepare saliva solution by spitting 3 ml of saliva into a clean beaker and diluting it with
3 ml of distilled water. (Remember to rinse your mouth first).
2. Place 1 ml of the saliva solution in each test tube labelled A1, B1, C1, D1 and E1.
3. Place 5 ml of 1% starch solution in another set of test tubes labeled A2, B2, C2, D2 and E2.
4. Place the two test-tubes labelled A1 and A2 into the water bath at 0 ºC (Figure 1). Leave
them for 5 minutes to stabilise the temperature.
5. Add the saliva solution from test-tube A1 into the 1% starch solution in test-tube A2 to start
the reaction. Immediately stir the mixture and start the stopwatch.
6. For every minute, take out a drop of solution from test-tube A2 and mix it with a drop of
iodine solution on a white tile to test the present of starch. (Figure 2)
7. Conduct iodine tests at a shorter interval (in every 30 seconds) when the reaction is nearing
completion.
8. Stop the iodine test when the mixture does not show any changes.
9. Record the time taken for the complete hydrolysis of starch at 0 ºC.
10. Repeat steps 4 to 9 for test tubes B2, C2, D2 and E2 in the water baths at the following
temperatures: 20 °C, 30 °C, 40 °C and 50 °C respectively.
(The temperature of the water must be kept constant by adding hot /cold water into it).
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11. Record the time taken for each complete hydrolysis in the table below.
1
12. Plot the graph of rate of reaction ( ) against the temperature (T °C)
t
Results
Rate of reaction
Temperature Time taken for complete hydrolysis, t 1
Test-tubes
(°C) (minute) (min−1)
t
A2 0
B2 20
C2 30
D2 40
E2 50
Discussion
1. From the graph, estimate the optimum temperature for amylase activity.
…………………………………………………………………………………………………….
2. Explain the changes to the enzyme molecules and the rate of reaction beyond optimal
temperature.
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
3. (a) Calculate the temperature coefficient, Q10, between 20 °C and 30 °C.
(b) Explain your answer in 3(a).
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
Conclusion
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
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Experiment 5
Title: Oxygen consumption during respiration in plant
Objective: To determine the oxygen consumption during germination of mung bean seeds
Learning Outcome:
(a) to use volumeter correctly;
(b) to calculate the amount oxygen consumption required by germinating seeds.
Introduction
Seeds contain food materials such as carbohydrate. When a seed germinates, the carbohydrate
undergoes a series of oxidation-reduction reactions to liberate the energy (ATP) required for the
embryo to grow into a seedling. At the same time, carbon dioxide is also produced. The
germination of seeds is a process of aerobic respiration, therefore, oxygen is consumed in the
process. This experiment quantifies the amount of oxygen consumed by germinating mung
bean seeds.

15 − 20 ungerminated (dry) mung bean seeds
5 − 10 germinating mung bean seeds (prepared by soaking in water for 24 hours)
Volumeter (readily assembled before class)
Glass beads
Non-absorbent cotton
9 − 10 pellets Potassium hydroxide (KOH) (Caustic agent)
Marker fluid in dropping bottle
Labelling sticker
Vaseline
Retort stand
Glass tube 3 mm diameter (internal) & the side arm 30 cm length
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Procedure
(Student are advise to carry out this experiment in pair/group)
Syringe
Add a small
drop of
marker fluid
here
KOH
KOH
Wool Wool
Glass bead
Germinated seed
Ungerminated seed
Dry sand
Figure 1: Volumeter
1. Prepare a set of apparatus/volumeter as shown in Figure 1

2. Label the tubes as A and B and then fill as follows:
Tube A: 5 − 10 germinating seeds.
Tube B: 5 − 10 ungerminated seeds.
(Student should expect that the seed level in tube A is higher than tube B)
3. Add enough glass beads into tube B to bring the total volume equal to that in tube A.
4. Pack cotton loosely into each tube to a thickness of about 1.5 cm above the seeds
(tube A) or beads (tube B).
5. Add 9 − 10 pellets of KOH on top of the cotton in the tubes.
Caution: KOH is a caustic agent, therefore handle it with care and do not touch with your
bare hands.
6. Insert the stopper-syringe-side arm pipette assembly in place. Apply vaseline to all joints.
7. Add a small drop of marker fluid to the end of each pipette.
8. Gently withdraw the plunger of each syringe to adjust the position of the marker lies which
shouldbetween 0.80 and 0.90 cm on the scale.
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9. Adjust the pipette to a horizontal position. Record the initial position of the marker
5 minutes after setting up the apparatus.
10. Run the experiment for 60 minutes. Record the reading at 10 minute intervals.
11. Tabulate your reading in Table 1.
12. Plot a graph of volume of oxygen consumption against time.
Results
1. Data tabulation
Initial position of marker for the germinating seeds:__________________
Initial position of marker for the ungerminated seeds:_________________
Table 1: Tabulation of results
Germinating seeds Ungerminated seeds
Time Position of the Cumulative Position of the

Cumulative
(min) marker in the oxygen volume marker in the
oxygen volume
side arm pipette consumed* side arm pipette
consumed (ml)
(cm) (ml) (cm)
10
20
30
40
50
60
*Note: Use formula 2

h to calculate oxygen volume. h is distance travelled by the marker.
Total volume of oxygen consumed by germinating seeds:______________

Total volume of oxygen consumed by ungerminated seeds:_____________
Discussion
1. What causes the change in volume?
…………………………………………………….………………………………………………
2. (a) What is the purpose of potassium hydroxide (KOH) pellets in this experiment?
…………………………………………………….………………………………………………
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(b) What would be the expected result if the experiment is performed without adding KOH
pellets to the tube? Why?
…………………………………………………….………………………………………………
…………………………………………………….………………………………………………
3. If 820 ml of oxygen is used to completely oxidise 1 g of glucose during seed germination,
how much glucose are consumed by 10 germinating mung bean seeds per hour?
4. State two physical factors that may affect the accuracy in measuring the oxygen
consumption in this experiment.
…………………………………………………….……………………………………................
…………………………………………………….……………………………………................
Conclusion
…………………………………………………….………………………………………………
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Experiment 6
Title: Separation of photosynthetic pigments using paper chromatography
Objective:
(a) To identify the different photosynthetic pigments found in chlorophyll
(b) To determine Rf value of pigments
Learning outcome:
(a) to know the presence of different pigments in chlorophyll;
(b) to understand that molecules of different sizes move at different rates on paper
chromatography.
Introduction
Green plants need light energy which is trapped by photosynthetic pigments to carry out
photosynthesis. Light of different wavelengths is trapped by different photosynthetic pigments.
Chlorophylls and carotenoids are the two main groups of photosynthetic pigments. Chlorophyll
a and chlorophyll b being the most common forms of chlorophyll absorb light in the blue and
the red parts of the spectrum. The carotenoids are often referred to as accessory pigments. They
include carotene and xanthophyll which absorb light from the blue part of the spectrum. Paper
chromatography is a method used to separate the various photosynthetic pigments from one
another. These pigments can be identified by their colours and Rf values.

80% acetone
“Cekur manis” or Red spinach leaves
Mortar and pestle
Muslin (cheese cloth)
Boiling tubes/large test tube with stopper (24 × 150 mm)
Solvent (1 part 80% acetone and 9 parts petroleum ether)
Ruler
Whatman no. 1 chromatography paper strip (20 × 130 mm)
Scissors
Pin
Dark paper/black sugar paper
Retort stand/test tube rack
Procedure
Switch off the fans while conducting the experiment.

1. Prepare the pigment extract by grinding up fresh leaves with 5 ml acetone using a mortar
and pestle.
2. Squeeze out a thick pigment extract using a cheese cloth.
3. Cut out one end of the chromatography strip to form a pointed tip. Rule a pencil line across
the strip of paper, 20 mm from the pointed tip.
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4. Transfer the pigment extract by spotting onto the centre of the pencil line of a
chromatography strip using the head of a small pin as a dropper. Let the spot dry.
Repeat the process 15 to 20 times over on the same spot. (Smaller spot gives better result)
5. Put enough solvent into a large test tube.
6 Hang the chromatography strip to the stopper using a pin/clip (pointed end down into the
solvent). Place the stopper to seal the tube (Figure 1). Make sure that the strip is in a
perfectly vertical position. Solvent level must be 1 cm from the pigment spot. Place the
tube on a rack or stand it using retort stand.
7. Let the solvent moves until it reaches near the pin/clip. Take out the chromatography strip
when the solvent front comes close to the top. Mark the solvent front with a pencil. Quickly
dry it and measure the distance between the pigment spot and the solvent front.
8. Observe and mark the positions of the separated pigments.
9. Measure the distance from the pigment spot to the leading edge of each pigment.
10. Cover the chromatography strip with a dark paper to protect the pigments.
11. Calculate the Rf value of each pigment using the formula:
Distance moved by pigment

Rf =
Distance moved by solvent
12. Record all the data in the Table 1 on the next page.
Stopper
Pin
Chromatography paper
Chlorophyll spot
Solvent
Figure 1: Assembling of chromatography apparatus
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Results
(i) Table 1
Pigment colours
Distance moved by pigments (mm)
Distance moved by solvent (mm)
Rf
(ii) Stick your chromatogram in the space provided below.
Discussion
1. Identify the pigments based on their colour.
…….................................................................................................................................................
…….................................................................................................................................................
…….................................................................................................................................................
…….................................................................................................................................................
…….................................................................................................................................................
2. What are the factors that affect the movement of pigment during chromatography?
…….................................................................................................................................................
…….................................................................................................................................................
…….................................................................................................................................................
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3. (a) Based on Table 1, name the pigment which move furthest. Explain your answer.
…….................................................................................................................................................
…….................................................................................................................................................
…….................................................................................................................................................
(b) Relate the Rf value to the molecular size of the pigment.
…….................................................................................................................................................
…….................................................................................................................................................
Conclusion
…….................................................................................................................................................
…….................................................................................................................................................
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Experiment 7
Title: Comparison between C3 and C4 leaves
Objective:
(a) To compare the cell organisation of the C3 and C4 leaves
(b) To relate the structure of the cells to their functions
Learning outcome:
(a) to describe the differences between C3 and C4 leaf structure;
(b) to understand that C4 plant is more efficient in photosynthesis compared to C3 plant.
Introduction
Photosynthesis is a process that converts carbon dioxide and water into organic compounds
using light energy. It is important to supply the carbon and energy sources for all living
organisms. The process occurs mainly in plants and in some bacteria and algae. Photosynthetic
organisms are called photoautotroph. The process begins when energy from light is absorbed by
chlorophylls. Photosynthesis involves a series of biochemical reaction called Calvin cycle.
Based on their adaptive mechanism for photosynthesis plants can be divided into three different
groups which are C3, C4 and CAM. This adaptation helps to reduce a wasteful process called
photorespiration that consumes part of the sugar produced during photosynthesis.

Light microscope
Prepared slide of transverse section of Zea mays leaf
Prepared slide of transverse section of Helianthus sp. leaf
Procedure
1. Examine the slides under low power objective lens.
(a) Students should observe that in the C3 leaf
− the palisade mesophyll cells exist as one basic layer below the upper epidermal
layer
− the spongy mesophyll cells with large spaces in between cells
− the bundle sheath cells are small
(b) While in the C4 leaf

− the mesophyll cells are arranged to form a ring around the bundle sheath cells
− the bundle sheath cells are large
2. Make labelled plan drawings for each of the leaf sections.
3. Observe the slides under high power objective lens.
4. Make labelled drawings for the cells, which you have observed. State the magnification of
the drawing.
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Results
1. Plan drawing
C3 leaf
Microscope magnification: .............................................
C4 leaf
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2. High power drawing
C3 leaf
C4 leaf
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Discussion
1. Name the cell where the Calvin cycle takes place in C3 and C4 plants.
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
2. Name the cell where atmospheric carbon fixation occurs in C3 and C4 plants.
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
3. What is the special characteristic of the bundle sheath cells in C4 leaf?
…………………………………………………………………………………………………….
4. Why photosynthesis in C4 plant is more efficient than that in C3 plant?
…………………………………………………………………………………………………….
Conclusion
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
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2.5 Experiment for Second Term
Experiment 8
Title: Dissection of the mammalian respiratory system
Objective: To examine the structures of the main internal organs involved in respiration
(larynx, trachea, lungs, diaphragm, and rib cage)
Learning outcome:
(a) to dissect rat and use the dissecting instruments;
(b) to display, draw and label the respiratory organs;
(c) to describe the function of lungs, trachea, diaphragm, rib cage, and intercostal muscles.
Introduction
Respiration is the uptake of oxygen and the release of carbon dioxide. Oxygen is used within
the cells during the breakdown of fuel, primarily glucose. The chemical process by which cells
break down fuel molecules using oxygen, producing carbon dioxide and releasing energy is
called aerobic cellular respiration. Respiring organisms obtain oxygen from their environment.
In order for an organism to move oxygen and carbon dioxide into and out of its body, it must
have a thin, moist and relatively large respiratory surface.
Different organisms use different organs for respiration such as cell (single-celled organisms)
skin (amphibians), trachea (most insects and some spiders), gill (large aquatic animals) and
lung (mammals). In humans and other mammals, air moves into the lungs through the nose or
mouth and airways of the respiratory system. The movement of air into and out the lungs is
called breathing, and gas exchange takes place in the alveoli. Alveoli provide a perfect place for
carbon dioxide and oxygen to diffuse between the air in the lungs and the blood.

Rat (Rabbit optional)
Dissecting instruments
Dissecting board
Magnifying glass × 10
Thread
Dissecting pins
Diethyl ether
Killing jar
Cotton balls
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Procedure
1. Kill a rat using diethyl ether swabbed with cotton balls in a killing jar.
2. Place the rat on dissecting board with the ventral surface uppermost. Pin all four legs to
stretch the body.
3. Make a mid-ventral incision through the skin and cut forward as far as the lower jaw and
then cut backwards to the anus.
4. By holding the skin with a pair of forceps, cut off the connective tissues between the skin
and the body wall as far as possible around the animal’s body. Pull back and pin the skin
(Figure 1).
Figure 1
5. Cut off the ventral and lateral thoracic walls to expose the thoracic cavity.
6. Identify the thymus gland and remove it.
7. Cut off muscles and tissues of the neck to expose the trachea and larynx.
8. Cut above the larynx. Cut off the connective tissues attached to the trachea.
9. Remove together the heart, lungs, trachea, esophagus and larynx.
10. Carefully separate the esophagus from the heart. Pin the heart, larynx, trachea and lungs to
the board.
11. Make a labelled drawing of the heart, lungs, trachea and larynx.
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Result:
Labelled drawing
Discussion
1. How does the rib cage function during gas exchange in this animal?
.........................................................................................................................................................
.........................................................................................................................................................
2. (a) Describe the appearance and characteristic of a diaphragm.
.........................................................................................................................................................
(b) What is the importance of this characteristic of the diaphragm in relation to its function
during gas exchange?
.........................................................................................................................................................
.........................................................................................................................................................
.........................................................................................................................................................
3. Write the correct sequence of air flow during inhalation.
.........................................................................................................................................................
.........................................................................................................................................................
Conclusion
.........................................................................................................................................................
.........................................................................................................................................................
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Experiment 9
Title: Dissection of the mammalian blood circulatory system
Objective: To examine the structures of the main internal organs involved in blood circulation
Learning Outcome:
(a) to dissect rat and use the dissecting instruments;
(b) to display, draw and label the blood circulation organs;
(c) to describe the function of heart, artery and vein.
Introduction
The circulatory system is responsible for the transport of body fluids. It is composed of two
systems, the blood system and the lymphatic system. The blood circulatory system includes the
heart, arteries, veins and capillaries. The arteries are those vessels which carry blood away from
the heart. Their walls are muscular to allow for expansion in response to the forceful pumping
of the heart. The veins are those vessels which return blood to the heart. They are much less
muscular than the arteries and are frequently transparent. Blood pressure in the veins is not as
intense as in the arteries. The capillaries are the microscopic blood vessels which connect the
arteries to the veins. It is in the capillary beds that the major function of the circulatory system
is accomplished. Through the walls of these tiny vessels, oxygen and nutrients pass into the
tissues and carbon dioxide and metabolic wastes pass from the tissues into the blood for
removal. Oxygenated blood is carried by the arteries and deoxygenated blood is carried by the
veins. Only the pulmonary artery and the pulmonary vein are different in this respect.

Dissecting board
Thread
Dissecting pins
Diethyl ether
Killing jar
Cotton balls
Procedure
1. Kill rat using diethyl ether swabbed with cotton balls in a killing jar.
stretch the body.
then cut backwards to the anus.
and the body wall as far as possible around the animal’s body. Pull back and pin the skin.
5. Open up the thorax by cutting along the dotted lines shown in Figure 1.
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Cut through wall of thorax

along this dotted line
Wall of thorax (ribs and

intercostals muscles)
Diaphragm
Xiphoid cartilage (xiphisternum)
Figure 1
6. Tie a thread to the xiphoid cartilage and pull it back to stretch the diaphragm (Figure 2).
Neck muscles
Clavicle
First rib
Pectoral muscle
Thymus gland
Pulmonary artery
Rib
Pulmonary vein
⎧ Left atrium
Heart ⎨ Intercostal muscle
⎩ Left ventricle Phrenic nerve
Muscles of diaphragm
Thread attached to
L: lung xiphoid cartilage
Figure 2
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7. Remove the thymus gland from the surface of heart to expose the heart.
8. Remove fat around the great vessels (arteries and vessel) in the neck region with a pair of
forceps.
9. Pull the heart to your left and identify the structures as shown in Figure 3.
Innominate artery
leading to right common
Pectoral muscle carotid artery
Left common
carotid artery
Left anterior vena cava
Right anterior
vena cava Systemic (aortic) arch
Left ventricle
Left atrium Left pulmonary artery
Left pulmonary vein
Posterior (inferior)
vena cava
Diaphragm
Xiphoid cartilage
Figure 3
10. Pull the heart to your right and identify the point where the venae cavae enter the right
atrium. Make a labelled drawing to show main blood vessels (Drawing 1).
11. Carefully trace the right anterior vena cava and innominate artery from the thorax up into
the neck. Remove all the muscle obscuring these vessels (Figure 4).
Neck muscles
Right external jugular vein Left jugular vein
Pectoral muscle beneath Pectoral muscle
which is the clavicle Left common carotid artery
Innominate artery Left anterior vena cava
Right anterior vena cava Cut wall of thorax
Systemic arch
Figure 4
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12. Remove the pectoral muscle and clavicle to reveal the origin of the subclavian and internal
jugular veins.
13. Trace the innominate artery and notice the origin of the right subclavian artery. The
innominate now continues forward as the right common carotid artery. Make a labelled
drawing to show innominate artery, the right subclavian artery and the right carotid artery
(Drawing 2).
14. Remove the heart and soak in hot boiling water. Make a longitudinal section and use a
magnifying glass to examine heart chambers and make a labelled drawing (Drawing 3).
Results
Drawing 1
Drawing 2
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Drawing 3
Discussion
1. What is the function of carotid artery?
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
2. Why does the colour of the vein appear darker than the artery?
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
3. State the sequence of blood flow in heart chambers.
………………………………………………………………………………………………….....
…………………………………………………………………………………………………….
Conclusion
…………………………………………………………………………………………………….
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Experiment 10
Title: Dissection of the mammalian urino-genital system
Objective:
(a) To identify the urinary system of the rat
(b) To identify the genital system of the rat
Learning outcome:
(a) to describe the urinary system of the rat;
(b) to describe the male and female genital systems of the rat;
(c) to differentiate the male and female genital systems of the rat.
Introduction
The urino-genital system of the mammals in general and in the rats in particular consists of the
urinary or excretory and genital (male and female reproductive) systems. The two systems are
conveniently discussed together because they originate from adjacent tissues during their
development and utilize common ducts to discharge their products out of the body. The major
structures of the urinary system are the kidneys, ureters, urinary bladder and urethra. The major
structures of the genital system of the male are the testes, sperm ducts, urethra and penis. In the
female, they are the ovaries, fallopian tubes, uterus, and vagina. The overall purpose of the
reproductive system is to produce egg and sperm cells and bring them to fertilization and
nourishing the developing embryo until birth. The urinary system helps to remove nitrogenous
wastes from protein metabolism and other harmful substances and eliminate them in the form
of urea in urine. This system also helps to maintain the internal environment of the body.

Dissecting board
Thread
Dissecting pins
Diethyl ether
Killing jar
Cotton balls
Procedure
1. Kill a rat using diethyl ether swabbed with cotton balls in a killing jar.
stretch the body. Observe the external morphology of the sexual features of the rat.
then cut backwards. For the male rat, cut the skin towards posterior through the penis and
in between scrotums. For the female rat, cut the skin through the vagina and anus.
and the body wall as far as possible around the animal’s body. Pull back and pin the skin.
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5. Open up the body wall in the abdominal region by making a small incision in the mid-line
and cutting through to the xiphoid cartilage. Continue cutting to the side of the last rib on
both sides of the body. For the male rat cut this wall through the penis and for the female
rat, through the vagina. Pin the walls on the board.
(At this point, students should be able to examine all organs of the abdomen in their
respective positions (Figure 1).)
Upper incisor
Lower incisor
Salivary gland
Xiphoid cartilage
Lobes of liver
Stomach
Spleen
Cut edge of body wall
Colon
Small intestine
Caecum
Urino-genital system
Figure 1
6. Remove the alimentary system by cutting off the esophagus (to the point nearest to the
diaphragm) and end of rectum.
7. Identify urinary bladder, prostate gland (male), preputial gland and clitoris (female).
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A. For the male rat

1. Cut the ventral wall of the scrotum to the apex and locate the following: penis,
epididymis on the testis, vas deferens, seminal vesicle, Cowper gland, prostate gland
and coagulating gland (Figure 2).
2. Remove fat bodies to locate the kidneys and ureters.
3. Locate the adrenal gland on each kidney.
4. Make a labelled drawing of male urino-genital system.
Kidney
Ureter
Seminal vesicle
Coagulating gland
Bladder
Prostate gland Cut pubis
Fat attached to testis Muscle
Urethra
Spermatic
Cowper gland
Fat
Preputial gland Vas deferens
Penis
Epididymis
Scrotal sac
Gubernaculum
Figure 2: Male rat
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B. For the female rat

1. Locate the following: a pair of ovaries attached by mesenteries, the oviduct
(or fallopian tube), right and left horns of the uterus which combined in the middle to
form the vagina (Figure 3). For pregnant female fetuses (in the uterus) can also be
detected (Figure 4).
2. Remove fat bodies to locate the kidneys and ureters.
3. Locate the adrenal gland on each kidney.
4. Make a labelled drawing of female urino-genital system.
Mammary gland
Ovarian & uterine

Artery & vein
Ovary
Uterus
Bladder
Mammary gland Vagina
Muscle
Urethra
Cut pubis
Preputial gland
Urethral opening
Vaginal opening
Anus Clitoris
Figure 3: Virgin female rat
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Adrenal gland
Fat
Ovary
Fallopian tube
Ureter
Foetus
Placenta
Uterus
Bladder
Vagina
Urethra
Vaginal opening
Preputial gland
Clitoris and Urethral Anus

opening
Figure 4: Pregnant female rat
Note: Students should examine both male and female rats.
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Results
(a) Draw the urino-genital system of the male rat.
(b) Draw the urino-genital system of the female rat.
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Discussion
1. What is the role of adrenal gland in relation to kidney function.
……………………………………………………………………………………….....................
……………………………………………………………………………………….....................
2. Cowper gland, prostate gland and coagulating gland are referred to as the ____________.
3. State the different role of urethra in the male and female rat.
……………………………………………………………………………………….....................
……………………………………………………………………………………….....................
Conclusion
……………………………………………………………………………………….....................
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Experiment 11
Title: Embryogenesis in animal
Objective: To study stages during embryogenesis in animal
Learning outcome:
(a) to describe stages during embryo development up to gastrulation;
(b) to characterise embryo at cleavage, blastula and gastrula stages;
(c) to identify cell layers that give rise to different organs.
Introduction
Embryogenesis is the process of embryo formation and development. Embryogenesis occurs
immediately after ovum fertilisation (zygote) and lasted for up to 8 weeks. During the process
zygote undergoes rapid mitotic division to form an embryo. Generally, embryogenesis occurs in
3 stages i.e. cleavage, blastulation and gastrulation. Cleavage of the zygote starts 12 hours after
fertilization to form a 2-cells stage. Within 72 hours the 2-cells stage divides to form a 4-cells
stage and an 8-cells stage. Cleavage process lasted when a morula is formed. The morula will
then undergo cells reorganization process to form a blastocyst. The blastocyst is composed of a
layer of trophoblast cells surrounding blastocoels cavity. The blastocoels cavity contains a
group of cells called an inner mass. During gastrulation, the inners mass cells divide to form
three germ layers referred to as the endoderm, mesoderm and ectoderm.

CD of embryogenesis
Computer
LCD projector
Projector screen
Procedure
1. Observe the displayed slides of embryo at different stages during development.
2. Draw the embryo at 4-celled, morula, blastocyst and gastrula stages in the correct boxes.
Label your drawings.
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Results
Drawings of 4-celled, morula, blastocyst and gastrula stages.
Title: 4-celled Title: Morula
Title: Blastocyst Title: Gastrula
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Discussion
1. Name the male and female reproductive cells.
…………………………………………………………………………………………………….
2. What is the function of trophoblast cell?
……………………………………………………………………………………………….……
3. What is the function of the inner cell mass?
…………………………………………………………………………………………….………
4. Name two organs developed from the following cell layers.

(i) Mesoderm
……………………………………………………………………………………….....................
(ii) Endoderm
…………………………………………………………………………………….…………........
(iii) Ectoderm
………………………………………………………………………………………...……........
Conclusion
………………………………………………………………………………………………….…
…………………………………………………………………………………………….………
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Experiment 12
Title: Examination of slides: kidney, liver and pancreas
Objective: To study the general anatomy of kidney, liver and pancreas
Learning outcome
(a) to identify the structures in kidney, liver and pancreas;
(b) to examine the basic cellular organizations of kidney, liver and pancreas.
Introduction
Organs are made of various types of tissues. Basically, there are four types of tissues found in
higher animals including man: epithelial, connective, muscular and nervous tissues. The study
of these tissues is called histology. In this class you will examine the basic cellular structures of
the kidney, liver and pancreas of adult mouse using prepared sections of organs which are
permanently mounted on glass microscope slides.

Prepared microscope slides of kidney
Prepared microscope slides of liver
Prepared microscope slides of pancreas
Light microscope
Magnifying glass
Procedures
1. Observe the general anatomy or an outline of the longitudinal section of the organ using a
magnifying glass. The overall outline of the respective organ can be observed properly.
2. Place the slide on the microscope stage.
(Caution: It is advisable to observe slides from lower to higher magnifications.)
3. Adjust correct light intensity using the diaphragm and do fine-focusing in order to get a
clear image.
4. Examine the detailed cellular structures of the respective organs using medium power
magnifications by identifying different types of cell and their arrangements. This can be
done in stepwise, first using 4/10× magnification and later 40/60×
Note: you must have noticed that at low power magnification, the field of view (area of
observation) is larger than that at higher magnifications. At higher magnifications, larger
image can be seen as the field of view becomes smaller or restricted.
Observe and examine the slides with these guidelines.
5. Kidney is surrounded by capsule and divided into outer cortex and inner medulla
(Figure 1 & 2). Nephron or filtration unit of the kidney consists of a dilated renal
corpuscle, proximal convoluted tubules, thin and thick limb of Henle’s loop and the distal
convoluted tubules which are connected to collecting ducts.
The renal corpuscle or Malphigian corpuscle consists of tuft of capillaries or the
glomerulus, surrounded by a double layer of epithelial capsule called Bowman’s capsule.
The outer and the inner layers of the capsule is made of simple squamous epithelial cells
(Figures 3 & 4). Between the two layers is the urinary space which receives the fluid
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filtered through the capillary wall and the visceral layer. Around the renal corpuscle, the
proximal convoluted tubules and the distal convoluted tubules (with bigger lumen without
brush border cells) can be seen. The walls of these tubules are made of simple cuboidal
epithelial cells (Figures 3 & 4). For convenience observe these structures in the cortical
area (cortex) of the kidney using 40/60×.
Renal cortex
Renal medulla
Renal pelvis
Ureter
Figure 1: Longitudinal section of a kidney
Cortical nephron
(a) Renal cortex

Juxtamedullary
nephron
Nephron tubule
Bowman’s
capsule
Glomerulus Renal medulla
(b)
Collecting duct
Blood flow
Figure 2: General organization of a kidney and

renal corpuscle andrelated structures
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Figure 3: Section of renal cortex showing proximal (P) and distal convoluted tubules(D)
and glomerulus (G) (10x magnification)
Space within Simple cuboidal

capsule epithelium
Space within
nephron tubule
Capillary
Renal
corpuscle
Nuclei of simple
squamous epithelial
cells
Figure 4: Section of renal cortex showing renal corpuscle, glomerulus (capillaries),

simple squamous epithelialand simple cuboidal epithelial cells (40x magnification)
Liver:
The liver is covered by a thin connective tissue capsule (or Glisson’s capsule) which
becomes thicker at the hilum, where the portal vein and hepatic artery enter the organ and
where the right and left hepatic ducts and lymphatic exit.
The basic structural component of the liver is the liver cells or hepatocytes. These are
polyhedral epithelial cells arranged in rows or interconnected plates in liver lobules thus
liver consists of many lobules (Figure 5). The lobule is formed of a polygonal mass of this
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tissue with portal space at the periphery and a central (or centrolobular) vein in the centre.
The portal space is surrounded by arteries, veins and bile ducts (Figure 6). In between the
rows of the hepatocytes are the capillaries or liver sinusoids and in between the hepatocytes
are the bile canaliculi (canaliculus).
Hepatocytes
Central
vein
Sinusoid
Figure 5: Liver section showing central (centrolobular)

vein surrounding by hepatocytes and
blood capillary or sinusoid (40x magnification)
Branch of
hepatic artery
Branch of
portal vein
Biliary duct
Figure 6: Liver section showing a portal space with

branch of portal vein, branch of hepatic artery
and bile duct or bile canaliculus (10x magnification)
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Pancreas:
This is a mixed exocrine-endocrine gland which produces digestive enzymes and
hormones.The hormones are synthesized in clusters of endocrine epithelial cells known as
islets of Langerhans (Figure 7). These are rounded clusters of cells found embedded within
the exocrine pancreatic tissue. Each islet is surrounded by acinar cells, and consists of
lightly-stained polygonal or rounded cells arranged in cords separated by a network of
blood capillaries (Figure 7). It is possible to recognize α (glucagon-producing) and β
(insulin-producing) cells (Figure 8).
Acinar cell
Blood
capillaries
Cell cords
Figure 7: A section of pancreas showing an islet of Langerhans surrounded by acinar cells

(40/60 × magnification)
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Blood
capillaries
α cells
β cells
Figure 8: Section of an islet of Langerhansshowing α, β cells and blood capillaries

(40/60 × magnification)
Results
1. Draw a plan diagram of a kidney. Label your drawing.
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2. Draw and label structures in renal cortex showing a renal corpuscle and its related
capillaries and tubule at medium power magnification. Indicate simple squamous and
cuboidal epithelial cells in the drawing.
Microscope magnification: …………………………….
3. Draw and label a diagram showing the central vein, blood cells, hepatocytes and sinusoid at
medium power magnification.
Microscope magnification: …………………………….
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4. Draw and label the portal area showing the following structures: branch of bile duct, branch
of hepatic artery, erythrocytes and branch of hepatic portal vein at medium power
magnification.
Microscope magnification: ……………………….
5. Draw and label diagram showing cells and structure of islet of Langerhans at medium
power magnification.
Microscope magnification: ………………………..
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Discussion
1. State the type of cell lining the tubules in Figure 2(a).
...…………………………………………………………………………………………………..
...…………………………………………………………………………………………………..
2. Where can you locate erythrocytes in the liver?
...…………………………………………………………………………………………………..
...…………………………………………………………………………………………………..
3. How do you differentiate between an artery and a vein in the liver?
...…………………………………………………………………………………………………..
...…………………………………………………………………………………………………..
4. Why is liver more compact than kidney?
...…………………………………………………………………………………………………..
...…………………………………………………………………………………………………..
Conclusion
...…………………………………………………………………………………………………..
...…………………………………………………………………………………………………..
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2.6 Experiment for Third Term
Experiment 13
Title: Classification of organisms using a dichotomous key
Objective: To classify organisms according their morphological characters
Learning outcome:
(a) to categorise organisms based on their morphological characters;
(b) to construct spider key;
(c) to construct a dichotomous key.
Introduction
A dichotomous key is a series of opposing descriptions that describe physical and anatomical
characteristics of different organisms. Biologists use dichotomous keys as a tool to identify
unknown plants and animals. Organisms are classified according to whether a particular
characteristic is present or absent. A choice between the two statements is made that best fits
the organism in question. In this experiment, you will construct a dichotomous key for a group
of organisms.

Light microscope
Magnifying glass
Amoeba/Paramecium slide
Hydra slide
Ant
Snail
Prawn
Marchantia/Funaria
Dryopteris
Grass
Procedure
1. You are given the following characteristics of organisms in Table 1 to assist you in
constructing a dichotomous key.
Table 1: Characteristics of organisms
with chlorophyll with exoskeleton
without chlorophyll without exoskeleton
produces spores with jointed legs
does not produce spores without jointed legs
has leaves (fronds) body divided into head, thorax and abdomen
has thalloid body body divided into cephalothorax and abdomen
unicellular organisms
multicellular organisms
2. Examine the external features of the organisms.
3. Match the features with the characteristics in Table 1.
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4. Group these organisms according to their contrasting features and construct a spider key.
5. Construct a dichotomous key.
Results:
1. Spider key
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2. Dichotomous key
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Discussion
1. (a) List down two features, which enable biologists to group ant and prawn into the same
phylum.
.........................................................................................................................................................
.........................................................................................................................................................
(b) Name the phylum.
.........................................................................................................................................................
2. State another structural difference to differentiate between Marchantia/Funaria and

Dryopteris.
Marchantia/Funaria........................................................................................................................
Dryopteris........................................................................................................................................
Conclusion
.........................................................................................................................................................
.........................................................................................................................................................
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Experiment 14
Title: Preservation techniques
Objective: To learn basic techniques for preserving animal and plant specimens
Learning outcome:
(a) to preserve animal and plant specimens using wet preservation techniques;
(b) to preserve animal and plant specimens using dry preservation techniques.
Introduction
Taxonomy is the study of classification of animals and plants based on their morphological
characteristics. Both fresh and live specimens can be used in this study. However, in many
instances, these specimens are normally not easy to handle or to maintain, prone to damages
and readily loose their important external features. As an alternative, there are techniques
available nowadays whereby fresh specimens can be preserved properly and their features
remained intact for a long time. Specimens can be preserved either in solution (wet preservation
technique) or in dry form (dry preservation technique). In this class you will learn basic
preservation techniques for specimens that are easily available around you.

Formalin acetic acid (FAA) solution
70% ethanol
Diethyl ether
Insect pins
Polystyrene board
Insect box
Labelling papers
50 ml test tubes
5 ml specimen bottles
50 ml specimen bottles
Petri dishes
Cotton wool or cotton balls
Newspapers
Magnifying glass
Drying oven
Live insects: house fly (small insect) and dragonfly (large insect)
Plant specimens: fresh fern leaf of Dryopteris and liverwort Marchantia/Funaria
Napthelene balls
Pencil
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Procedures
Part A: Wet preservation technique for house fly
1. Wet a cotton ball with diethyl ether and place it at the bottom of a 50 ml test tube.
2. Put a few house flies into the test tube and stopper the test tube for 1-2 minutes for fresh
killing of the specimen.
3. Draw out the dead house flies and place them into a 5 ml specimen bottle containing 70%
ethanol for preservation.
4. Label this specimen with a labeling paper with the following information:
− Name of specimen
− Date of collection
− Habitat
− Name of the collector
5. Place the labeling paper into the specimen bottle.
Part B: Dry preservation technique for house fly

1. Wet a cotton ball with diethyl ether and place it at the bottom of a 50 ml test tube.
2. Put a few house flies into the test tube and stopper the test tube for 1-2 minutes for fresh
killing of the specimen.
3. Carefully, pierce an insect pin through the thorax of the fly (Figure 1).
4. Place the dead flies in a Petri dish and dry them in an oven at 40 − 50 ºC for 24 hours.
Pin
Figure 1: House fly
5. Stand the fly on a polystyrene board.

− Habitat
7. Continue drying this specimen in the oven (if necessary).
8. Keep the dried specimen in an insect box provided with naphthalene balls.
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Part C: Dry preservation technique for dragonfly

(Note: The wings of a dragonfly are quite long and delicate and folding them may cause some
damages. Therefore, dragonfly specimens are usually preserved in a dry form instead of the wet
form)
1. Hold a live dragonfly and observe the ventral side.
2. Using your thumb and index finger, hold the live dragonfly by the sides of its thorax and
squeeze it gently for 1 to two minutes (or until death).
3. Straighten the wings horizontally (care must be taken that straightening must be done
immediately after step 2 before the body and the wings become hardened).
Note: For larger insect such as Locust, cut the abdomen dorsally to remove the digestive
system to prevent the insect from rotting and to ensure good quality of preservation
4. Pierce an insect pin through the thorax and stand the specimen on a polystyrene board
(Figure 2).
Pin
Figure 2: Dragonfly
5. Place the specimen in the oven for drying for a maximum of 48 hours (it is important to
make note on the colour of the body before drying).
6. After drying, label this specimen with a labeling paper with the following information:
− Habitat
7. Keep the dried specimen in an insect box provided with naphthalene balls.
Part D: Wet preservation technique for Marchanti /Funaria

1. Fill a 50 ml specimen bottle with FAA solution.
2. Immerse Marchantia/Funaria specimen into the bottle and make sure it is fully submerged
in FAA solution. Note the colour of the specimen.
− Habitat
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Part E: Dry preservation technique for Dryopteris leaf

1. Place a Dryopteris leaf between two pieces of newspapers and press it with a heavy object
on a flat surface to flatten it.
2. Leave it to dry at room temperature for a week.
3. Label this specimen with a labeling paper with the follong information:
− Habitat
Results
Preserved house fly, dragonfly, Marchantia, Dryopteris
Discussion
1. Describe the use of 70% ethanol
…………………………………………………………………………………………………….
2. (a) Why is it necessary to take note on the color of specimen before drying or before
soaking it in FAA solution?
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
(b) Why squeezing the thorax will kill the insect?
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
Conclusion
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
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Experiment 15
Title: Collections of 10 different insect orders and 10 different plant families
Objective:
(a) To learn methods of collecting insects and plants
(b) To learn methods of preserving insect and plant specimens
(c) To strengthen the theoretical understanding of naming and classification of organisms
Learning Outcome:
(a) to collect insect and plant specimens;
(b) to preserve insect and plant specimens;
(c) to classify the insect and plant specimens under different order and families.
Introduction
Insects and plants are major biotic components in an ecosystem. They can be found easily in
terrestrial and aquatic habitats. Many insects and plants coexist with humans in harmony. There
are more than 30 orders of insects and more than 300 families of plants in the world. Malaysia
is one of the 17 megadiversity countries, and the major part of the biodiversity in Malaysia
comes from insects and plants. In this class you will learn sampling and preservation methods
of some Malaysian local insect and plant specimens.
For insect For plant

Insect pooter or aspirator Plastic bags
Insect net Thread
Collecting tubes Old newspapers
Killing jar Plant press
Magnifying glass Measuring tape
4 − 10% formalin Plant cutter
Tray Herbarium paper (Drawing block)
White paper Needle
Insect pin
Insect box
Naphthalene balls
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Procedure
(Note: For this experiment students should work in a group of 3 to 5)
A. Collecting insects
1. Insect collection must consist of at least 10 different orders.

2. Some of the methods that can be used to collect insects are as follows:
(i) Beat small trees and tree branches using a sweeping net (sweep netting). This causes
the insects to fall into the net. Remove the insects, place them on a white paper and
examine them.
(ii) Hold a beating tray underneath small trees or tree branches, shake or beat the trees.
Insects will fall onto the tray and examine them.
(iii) Butterfly net is used to catch butterflies and other flying insects.
(iv) Use a light trap or span a white material vertically and also place another one at the
base. Switch on a light bulb. This will attract insects, which come out at night. The
insects can then be collected and examine.
(v) Use an aspirator or pooter to catch small insects through suction.
3. Transfer the collected insects into collecting tubes or killing jar.
4. Preserve the insects following the correct procedures according to their sizes.
5. Clearly display the collected insects. Label each insect as follows:
Local name: ……………………………………………

Order: …………………………………………………..
Location: ……………………………………………….
Habitat: …………………………………………………
Date of collection: ……………………………………..
Collector’s name: ………………………………………
Date: ……………………………………………………
Note: Please refer Mohamed Salleh (1990). Pengumpulan, Pengawetan dan Pengelasan
Serangga. DBP, Kuala Lumpur for further details.
Solomon, Berg & Martin (2011). Biology (9th edition) , Brooks/Cole Cengage Learning
B. Collecting plants
1. Plant collection must consist of at least 10 different families.
2. Plant specimens collected must be normal that is
(i) not abnormal
(ii) not too young
(iii) not too old
3. Press specimens immediately after collection. Only the leaf and stem (flowers if any) need
to be preserved (Figure 1).
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Figure 1: An example of plant press
4. Use pieces of paper 6 in × 10 in or 13 in × 8 in to display the specimens. Mounts the

specimen on herbarium paper by stitching the specimen with cotton thread.
(Note: Use of exercise books to replace loose pieces of paper is not allowed.)
5. Label each plant specimen as follows:
Local name: ……………………………………………

Family: …………………………………………………..
Location: ……………………………………………….
Habitat: …………………………………………………
Date of collection: ……………………………………..
Collector’s name: ………………………………………
Date: ……………………………………………………
6. Place the plant specimens in protective plastic jackets and displayed them in ring folders.
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Experiment 16
Title: Ecological study of a terrestrial habitat
Objective:
(a) To determine plant distribution in a terrestrial habitat
(b) To study the biotic and abiotic factors in a terrestrial habitat
Learning Outcome
(a) to use the correct technique of quadrat sampling;
(b) to estimate plant distribution in a terrestrial habitat;
(c) to relate soil pH and plant distribution in a terrestrial habitat.
Introduction
Plants and animals are biotic factors that play an important role in ecosystem. The population of
plants and animals in a particular habitat can be measured by methods of quadrat and line
transect. The samples obtained from quadrats are representative of the habitat in general.
Abiotic factors such as pH also play a big role in the distribution of plants in a particular
habitat. In this study your will learn a quadrat sampling technique for estimation of plant
population in a chosen terrestrial area.

1m × 1m quadrat
5 test-tubes with stopper
Test-tube rack
Spatula
10 cm3 pipette
Universal indicator
Barium sulphate powder
pH chart
Soil borer
5 plastic bags
Marker pen
Procedure
A. Determination of plant population using a quadrat sampling technique
1. Identify 5 dominant plant species (common/scientific name) in the selected habitat.
2. Randomly throw in the quadrat.
3. For each quadrat,
(a) identify the presence of the individual plant species. Record your data in Table 1;
(b) count the number of the individual plant species. Record your data in Table 2;
(c) calculate the area of coverage of the individual plant species. Record your data in
Table 3.
4. Repeat steps 2 and 3 for the next 9 quadrats.
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5. Attach photos of the following:

(a) sampling activity in the field
(b) quadrat sampling
(c) plant species observed in the quadrat
Formula:
(a)
(b)
(c)
(d)
(e)
(f)
A. Determination of soil pH
(Note: Soil samples are obtained from 5 quadrats above)
1. Collect the soil samples by pressing the soil borer into the soil.
2. Use the piston to remove the soil sample from the borer.
3. Put one full spatula of barium sulphate into a dry test tube.
4. Add one full spatula of soil into the test tube.
5. Fill the test tube with distilled water until ¾ full.
6. Add 5 − 6 drops of Universal indicator.
7. Place a stopper and shake the tube well. Allow the tube to stand for short while until
a clear coloured liquid formed at the top.
8. Compare the colour of the liquid with the pH chart and record the pH.
9. Repeat steps 3 to 8 for the next four soil samples.
10. Record all pH readings in Table 4.
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Results
Table 1: Presence of plant species
Key:
A: ………………………………………………………………………………………………………
B: ………………………………………………………………………………………………………
C: ………………………………………………………………………………………………………
D: ………………………………………………………………………………………………………
E: ………………………………………………………………………………………………………
Quadrat Total
Relative
number of Species
Plant species
quadrat frequency
species 1 2 3 4 5 6 7 8 9 10 frequency
containing (%)
(%)
species x
Total
Table 2: Number of plant species
Quadrat Total Relative

Species
Plant number species
density
species 1 2 3 4 5 6 7 8 9 10 of density
(m2)
species x (%)
Total
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Table 3: Area of coverage
Quadrat Total area

Relative
of Species
Plant species
coverage coverage
species 1 2 3 4 5 6 7 8 9 10 coverage
of species (%)
(%)
x
Total
Table 4: pH of the soil sample
Soil sample Colour pH
Average pH
Discussion
1. State the purpose of using barium sulphate.
…………………………………………………………………………………….........................
2. State the relationship between the soil pH and the most abundant plant in the habitat.
…………………………………………………………………………………….........................
Conclusion
…………………………………………………………………………………….........................
…………………………………………………………………………………….........................
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Experiment 17
Title: Mendelian inheritance
Objective: To test whether the results of monohybrid and dihybrid crosses in corn obey
Mendel’s Laws using χ2 test
Learning outcome:
(a) to identify the different phenotypes in monohybrid and dihybrid crosses in corn;
(b) to use χ2 to test the hypothesis of monohybrid and dihybrid crosses.
Introduction
The basic rules governing inheritance were first discovered by Gregor Mendel. Genes
controlling characteristics in organisms are inheritable. Alleles are alternative forms of the gene
for one characteristic. If only a pair of alleles (for one characteristic) is involved, the cross
determining of the characteristic is known as a monohybrid cross. If two pairs of alleles (for
two different characteristics) are involved, the cross is known as a dihybrid cross. Genetic
constitution (all genes/alleles present) of an individual is known as genotype. On the other
hand, physical appearance of an individual as a product of expression of alleles is known as
phenotype.
The ratios 3:1 and 9:3:3:1 are hypothetical estimations of phenotype classes from monohybrid
and dihybrid crosses respectively according to Mendelian inheritance. The χ2 test can be used
to determine if the ratio of a cross deviates from the expected ratio. In other words, χ2 test is
used to test if a cross follows Mendel’s Law. The χ2 test takes into account the sample size and
the number of parameters (degree of freedom). In Mendelian inheritance test, the degree of
freedom is one less than the number of phenotype classes.
1. Ears of corns from monohybrid crosses

(a) The phenotypes: purple and yellow seeds
(b) The phenotypes: smooth and wrinkled seeds
2. Ears of corns from dihybrid crosses

The phenotypes: purple / yellow and smooth / wrinkled seeds.
3. Pins
χ2 table
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Table 1: χ2 table
n
p .99 .98 .95 .90 .80 .70 .50 .30 .20 .10 .05 .02 .01
1 .00016 .00063 .0039 .016 .064 .148 .455 1.074 1.642 2.706 3.841 5.412 6.635
2 .0201 .0404 .103 .211 .446 .713 1.386 2.408 3.219 4.605 5.991 7.824 9.210
3 .115 .185 .352 .584 1.005 1.424 2.366 3.665 4.642 6.251 7.815 9.837 11.341
4 .297 .429 .711 1.064 1.649 2.195 3.357 4.878 5.989 7.779 9.488 11.668 13.277
5 .554 .752 1.145 1.610 2.343 3.000 4.351 6.064 7.289 9.236 11.070 13.388 15.086
6 .872 1.134 1.635 2.204 3.070 3.828 5.348 7.231 8.558 10.645 12.592 15.033 16.812
7 1.239 1.564 2.167 2.833 3.822 4.671 6.346 8.383 9.803 12.017 14.067 16.622 18.475
8 1.646 2.032 2.733 3.490 4.594 5.527 7.344 9.524 11.030 13.362 15.507 18.168 20.090
9 2.088 2.532 3.325 4.168 5.380 6.393 8.343 10.656 12.242 14.684 16.919 19.679 21.666
10 2.558 3.059 3.940 4.865 6.179 7.267 9.342 11.781 13.442 15.987 18.307 21.161 23.209
p − Probability
n − Degree of freedom
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Procedure
Part A: Monohybrid cross
1. Examine and record the phenotypes of the seeds on the ear of corn from monohybrid cross
given in Table 2.
2. Count all the seeds on the ear according to phenotypes and record your observed data. (You
may use pins to assist you in counting)
Caution: Pins should be inserted in between the rows but not on the seed.
3. Calculate and record the expected number of each phenotype according to ratio 3:1.
4. Calculate the χ2 value and test the significance of deviation of the observation from the
expectation at 95% confident level.
Results
Table 2: Monohybrid cross
Phenotype Expected Observed Expected number Divergence (o − e)2 / e

ratio number of count (o − e)
(o) (e)
Total χ2 =
Discussion
1. What is the degree of freedom for the monohybrid cross?
…………………………………………………….………………………………………………
2. State the dominant phenotype and the recessive phenotype.
…………………………………………………….………………………………………………
…………………………………………………….………………………………………………
3. What is the χ2 value for this degree of freedom at 95% confidence level?
…………………………………………………….………………………………………………
Conclusion
…………………………………………………….………………………………………………
…………………………………………………….………………………………………………
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Part B: Dihybrid cross
Procedure
1. Examine and record the phenotypes of the seeds on the ear of corn from dihybrid cross
given in Table 3.
2. Count all the seeds on the ear according to phenotypes and record your observed data.
(You may use pins to assist you in counting)
Caution: Pins should be inserted in between the rows but not on the seed.
3. Calculate and record the expected number of each phenotype according to ratio 9:3:3:1.
4. Calculate the χ2 value and test the significance of deviation of the observation from the
expectation at 95% confident level.
Results
Table 3: Dihybrid cross
Phenotype Expected Observed Expected number Divergence (o − e)2 / e

ratio number of count (o − e)
(o) (e)
Total χ2 =
1. What is the degree of freedom for the dihybrid cross?
…………………………………………………….………………………………………………
2. What is the χ2 value for this degree of freedom at 95% confidence level?
…………………………………………………….………………………………………………
Conclusion
…………………………………………………….………………………………………………
…………………………………………………….………………………………………………
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Experiment 18
Title: Extraction of plant DNA
Objective: To extract DNA from onion cells
Learning outcome:
(a) to indicate the presence of DNA in living cells;
(b) to describe the physical characteristics of DNA.
Introduction
Deoxyribonucleic acid (DNA) is a nucleic acid that contains the entire genetic information of
an organism. DNA is formed from nucleotide subunits namely thymine (T), adenine (A),
guanine (G) and cytosine (C). In a cell, DNA exists in the form of double-stranded helix
molecule due to pairing between A and T, and G and C. These two strands run in opposite
directions to each other and are therefore, anti-parallel. A segment of DNA that carries the
genetic information is called a gene. Eukaryotic cells store their DNA in organelles while
prokaryotic cells store their DNA in the cytoplasm.

Ice cold isopropanol
Onion (Indian onion gives better result)
DNA extraction kit (which contains homogenising sticks, extraction buffer, conical
centrifugation tubes, plastic pipette)
Knife
Procedure
1. Peel one onion bulb and cut it into small pieces.
2. Transfer the onion into an empty conical centrifugation tube.
3. Using a plastic pipette, transfer 3 ml of extraction buffer into the tube.
4. Gently homogenise the onion by using the homogenising stick.
(Caution: Vigorous movement will cause formation of excessive bubbles and reduce the
volume of extraction buffer.)
5. Transfer 2 ml of aqueous phase (from step 4) into a new conical tube.
6. Add 2 ml of ice cold isopropanol into the tube.
7. By slanting the tube, gently mix the aqueous and organic phases with a new homogenising
stick in a circular motion.
8. Remove the homogenising stick and observe the DNA at end of the stick.
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Results
1. Draw the appearance of DNA on the stick.
2. Describe the physical characteristics of DNA.
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
Discussion
1. Where is the location of DNA in a cell?
…………………………………………………………………………………………………….
2. What is the purpose of homogenising stage?
…………………………………………………………………………………………….………
3. What is the function of isopropanol in this experiment?
…………………………………………………………………………………………….………
4. What method could be used to visualise the presence of DNA in a solution?
……………………………………………………………………………….……………………
5. What is the basic unit of DNA?
……………………………………………………………………………….……………………
6. When does the DNA replicate in a cell cycle?
……………………………………………………………………………….……………………
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Conclusion
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
…………………………………………………………………………………………………….
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MAJLIS PEPERIKSAAN MALAYSIA

This manual consists of 125 printed pages.

Part 1: Teacher’s Manual for Biology Coursework

1.2 General Information 1

1.3 Recording of Assessment Marks 2

1.5 Practical Work Assessment Guide 4

1.6 Title of the experiment 6

1.7 Table of Summary of Experiment 7

1.8 Marking scheme/Teacher guide 8

1.9 List of Apparatus and Materials 32

Part 2: Student’s Manual for Biology Coursework

2.2 Assessment of Practical Work 46

2.3 Title of the experiment 48

2.4 Experiment for First Term 49

2.5 Experiment for Second Term 75

2.6 Experiment for Third Term 102

BIOLOGY COURSEWORK STPM 2016

Part 1: Teacher's Manual for Biology Coursework

1.2 General Information

1.2.4 Experiments are to be carried out either individually or in groups as recommended in

BIOLOGY COURSEWORK STPM 2016

1.3 Recording of assessment marks

BIOLOGY COURSEWORK STPM 2016

BIOLOGY COURSEWORK STPM 2016

1.5 Practical Work Assessment Guide

Microscope Manipulative • Skill in handling microscope

Result • Skills in drawing/accuracy, labels,

Discussion • Ability to answer questions correctly 2–6

Conclusion • Ability to make relevance conclusion 1–2

Biochemistry Manipulative • Preparation of materials, procedures,

Result • Ability to label and plot a graph

Discussion • Ability to answer questions correctly 5–8

Conclusion • Ability to make relevance conclusion 1–2

Dissection Manipulative • Ability to follow instructions on how to

Result • Skill in drawing and labels the organ

Discussion • Ability to answer questions correctly 5–6

Conclusion • Ability to make relevance conclusion 1–2

BIOLOGY COURSEWORK STPM 2016

Ecology Manipulative • Skill in collection and preservation of

Result • Skill in labeling and displaying

Discussion • Ability to answer questions correctly 2–4

Conclusion • Ability to make relevance conclusion 1–2

Genetic Manipulative • Ability to follow instruction

Result • Perform correct calculations

Discussion • Ability to answer questions correctly 6

Conclusion • Ability to make relevance conclusion 1–2

BIOLOGY COURSEWORK STPM 2016

1.6 Titles of Experiments

A Compulsory 1; 2; 7; 11; 12 3; 4; 5; 6 8; 9; 10 13; 14; 15; 16 17; 18

B Compulsory 1; 2; 11; 12 4; 5; 6 8; 9; 10 13; 15; 16 17; 18

BIOLOGY COURSEWORK STPM 2016

1.8 Marking Scheme/Teacher’s Guide for Experiments

Conclusion (Suggested) (D)

BIOLOGY COURSEWORK STPM 2016

Conclusion (suggested) (D)

BIOLOGY COURSEWORK STPM 2016

Sucrose concentrations (M) 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

− Correct volume of stock solution and distilled water used as shown

(b) Tabulation result

Test tube 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

(c) Plotting of graphs

BIOLOGY COURSEWORK STPM 2016