8

Research 2 Activity Sheet

Quarter 3 – MELC 5
Week 5
Laboratory Techniques and
Methods

REGION VI – WESTERN VISAYAS

Research 2
Activity Sheet No. 4- Laboratory Techniques and Methods
First Edition, 2021

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Introductory Message
Welcome to Research II!

The Learning Activity Sheet is a product of the collaborative efforts

of the Schools Division of San Carlos City and DepEd Regional Office VI -
Western Visayas through the Curriculum and Learning Management
Division (CLMD). This is developed to guide the learning facilitators
(teachers, parents, and responsible adults) in helping the learners meet the
standards set by the K to 12 Basic Education Curriculum.

The Learning Activity Sheet is self-directed instructional materials

aimed to guide the learners in accomplishing activities at their own pace and
time using the contextualized resources in the community. This will also
assist the learners in acquiring lifelong learning skills, knowledge, and
attitudes for productivity and employment.

For learning facilitator:

The Research 2 Activity Sheet will help you facilitate the leaching-learning
activities specified in each Most Essential Learning Competency (MELC) with
minimal or no face-to-face encounter between you and the learner. This will be
made available to the learners with the references/links to ease independent
learning.

For the learner:

The Research 2 Activity Sheet is developed to help you continue learning

even if you are not in school. This learning material provides you with meaningful
and engaging activities for independent learning. Being an active learner, carefully
read and understand the instructions then perform the activities and answer the
assessments. This will be returned to your facilitator on the agreed schedule.
 Name of Learner: ________________________________________________________

Grade and Section: ___________________________ Date:_____________________

RESEARCH 2 ACTIVITY SHEET NO. 4

Laboratory Techniques and Methods

I. Learning Competency with Code

o Identify the standard methods and techniques used in performing
experiments (SSP_RS8-SA-III-o-q17).

II. Background Information for Learners

In making scientific research, designing and conducting an experiment is of
great importance. An experiment is a set of procedures that are used by the
investigator to be able to collect the necessary information that is needed to verify
or test the truthfulness of the hypothesis or to answer the research questions. In
an experiment, all the essential conditions are controlled so that the results can
only be attributed to the independent variable (Lahorra, 2020).

Experiments are done under controlled conditions so that the results can be
achieved. Important concepts must be considered in learning about designing and
conducting experiments.

Study the table below.

Set-up of the Experiment

Concepts Experimental Control + Control -
Controlled variables ▪ Kind of tomato ▪ Kind of ▪ Kind of
are factors that are plant tomato plant tomato plant
maintained in the ▪ Type of soil ▪ Type of soil ▪ Type of soil
experiment. This is ▪ Amount of ▪ Amount of ▪ Amount of
done to avoid other water water water
factors intervening in ▪ Amount of ▪ Amount of ▪ Amount of
the results. sunlight sunlight sunlight

A dependent variable ▪ Growth of ▪ Growth of ▪ Growth of

is a factor or pechay plants pechay plants pechay plants
condition that might in terms of in terms of in terms of
be altered. The effect height (cm) height (cm) height (cm)
could be observed
and measured.

An independent ▪ Commercial ▪ Without

variable is a factor or ▪ Organic fertilizer fertilizer
condition that might fertilizer
be affected. The
effect could be
observed and
measured.
Experimental unit If a researcher wants to find out the growth rate of
refers to the subject pechay plants after three weeks wherein they will receive
or object which is different treatments (with organic fertilizer and without
introduced with a fertilizer). Therefore, the pechay plants are the
stimulus and is then experimental unit.
measured.
Treatment is the Using the same example on the experimental unit, the
stimulus introduced treatments introduced are – (a) adding organic fertilizer on
to the experimental the soil and (b) without adding fertilizer on the soil.
subjects.
Repeated Take for example if researchers wanted to test a certain
measurements are medication on patients with Diabetes II, they will be first
the measurements tested to measure their glucose level before they are given
that are made on a specific treatment. After the treatment has been
each subject over implemented, patients will be tested again and they might
several periods in undergo nutritional counseling as well.
some experiments.
Repeated Using the same example above, the measurement
measurement design design to be used is a crossover design – “two-period,
refers to the design two-treatment” which means the research participants
which includes are randomly assigned to receive two different
repeated treatments: either A then B, or B then A.
measurements on
research where a
single treatment is
administered to a Then also applicable to research studies that may
subject or it involves involve multiple factors affecting the results gathered
designs of treatment from it.
that are modified
before each
measurement.

The purpose of the control group or control treatment is for comparison. The
results from the experimental treatment will be compared to the results of the
control group. This helps in making valid conclusions about how organic fertilizer
showed its effects on the growth of the plants. There are two types of control groups
in experiments: negative and positive control groups. A negative control group
receives a negative control treatment (this is without treatment or a treatment that
will not give an effect). A positive control group receives a positive control
treatment, a treatment that is already known for a particular effect.

LABORATORY TECHNIQUES AND METHODS

Aside from doing the Scientific Method as a general method of conducting any
Science-related activities, laboratory methods and techniques are essentially
introduced since experiments are mostly done in laboratories. Cambridge
dictionary defines a laboratory as “a room or building with scientific equipment
for doing scientific tests for teaching Science, or a place where chemicals or
medicines are produced.” On the other hand, laboratory techniques are
procedures and practices that are required in utilizing various equipment inside
the laboratory.
 Here are some of the laboratory techniques and methods that you need to learn
as you conduct your research study:

A. Microbiological Techniques
In microbiology, microorganisms like archaea, bacteria, fungi, protozoa, viruses,
etc. are being studied.

The common microbiological techniques are discussed in detail below:

1) Sterilization
Microbiological media, a medium for growing bacteria or any other
microorganisms, must be sterilized by heating it in an autoclave (like a pressure
cooker) at 121°C for 15 minutes. This will kill all living organisms, i.e., spores.

2) Aseptic Techniques
This is done to lessen the chance of bacterial contamination. Furthermore, this
also involves disinfecting working area 4exposed to microbiological media to
minimize the possible contact by bacteria from the air and use flames to kill
bacteria that might enter the vessels after opening them.

3) Inoculation
This is the process by which bacteria are introduced to a media in various
means. For example (bacteria), from a drop in a heat-sterilized loop are dispersed
on the surface of the agar. This is the same with broth cultures. For bacteria in a
liquid, a sterile pipette is introduced to a Petri dish before the agar is poured on
the top (“pour plates”).

4) Incubation
This is the process of maintaining favorable conditions for growth and
development, i.e., microbiological culture.

The Petri dishes that contain the agar or tubes contain broth are incubated, for
example, a special apparatus is placed in a room with a fixed temperature
(usually at 37 °C, for pathogens, while for bacteria is 35 °C from the environment).
In school, incubation temperatures are set lower to minimize the growth of
potential pathogens.

B. Chemical
1) Spectrophotometry
This is a method by which you measure the amount of absorption and
transmittance of light. This is used widely for qualitative analysis in various fields
of Sciences (e.g., biochemistry, biology, chemistry, material and chemical
engineering, clinical applications, industrial applications, etc. ). A
spectrophotometer is a device that measures the number of photons (the intensity
of light) passing through a sample solution. This consists of two devices: a
spectrometer and a photometer. A spectrometer is a device that disperses and
 measures light. On the other hand, a photometer consists of a photoelectric
detector that measures the intensity of light.

2) Extraction

This is the conversion of compound/s from a solid or liquid state into another
phase or state.

Preparation of Crude Extracts from Plants

Studies about the medicinal properties of plants require the extraction of

crude compounds from plant parts such as leaves, flowers, and roots. Knowledge
on how to do the extraction process is necessary.

Step 1: Air Drying of Samples. After collecting the adequate amount of plant
samples, wash with clean water and dry5 at room temperature. The efficiency of
extracting crude compounds can be achieved when the plant samples are air-
dried.

Step 2: Grinding the Dried Samples into Powder. The purpose of this step is to
turn samples into small pieces (powder) using a Wiley Mill or if you do not have
one, you can use a pair of scissors. Compounds can easily dissolve in the solvent
when the plant sample is powdered.

Step 3: Soaking the Ground Plant Samples in a Solvent. Soaking the ground
samples in a specific solvent will make the compounds go to the solvent or will
dissolve in the solvent. This is already the start of the extraction process. The
kind of solvent you want to use depends on your study particularly on the
specific compound you want to extract from the plants. Most commonly used are
ethanol, acetone, petroleum ether, and hexane.

Step 4: Collecting the filtrate. The dissolved compounds from the plant samples
can be found in the filtrate, the liquid mixture. Filter the mixture using a filter
paper (usually Whatman #1 filter paper or as required by the procedure).

Step 5: Evaporating the solvent. This is the last step in preparing the crude
compounds of crude extract from the plant sample. You can evaporate the
solvent using a Rotatory Evaporator (Rotavap) or you can simplify by evaporating
the solvent by heating using an alcohol lamp. The crude extract should be in
semi-solid form.

Step 6: Recover the extract from the rotary evaporator container. The extract
should be semi-solid or sticky in appearance to ensure that the solvent ethanol
or other solvent chemicals are thoroughly removed.

3) Titration
This is the process of determining the concentration of a solution. Medwick &
Kirschner (2010) defined titration as “a common laboratory method of quantitative
 chemical analysis that used to determine the unknown concentration of an
identified analyte.” The common types of titration are acid-base titration and
redox titration. An acid-base titration depends on the neutralization between an
acid and a base when mixed in a solution. When there is a change in color in the
acid-base indicator signals the endpoint of the titration.

C. Physical

1) Aeration
This is the interchange of various gases (CO2 & O2) between the atmosphere and
soil and the various reactions that either consume or produce gases in the soil.

For example, the oxygen gas (O2) moves from the atmosphere to soil and is
consumed by plant roots and microorganisms. While carbon dioxide (CO 2) moves
from soil, where it is produced by the plant and microbial respiration, to the
atmosphere.
2) Centrifugation
This is the process of separating substances applying centrifugal force through
the use of a centrifuge. A centrifuge is a device used to separate particles or
macromolecules (i.e., cells, nucleic acids, proteins, and sub-cellular components).

3) Chromatography
The word chromatography means “color writing”, which chemists used to test
liquid mixtures. This is the process of separating organic and inorganic
compounds used by scientists to analyze and study these compounds.

There are four main types of chromatography: Gas Chromatography, Liquid

Chromatography, Paper Chromatography, and Thin-layer Chromatography.

The table below presents a detailed comparison of these types.

Types of Chromatography Description Application(s)

o Detects bombs in
- Used to analyze volatile airports
gases. o Identifies and
quantifies drugs, i.e.,
in alcohols
a) Gas Chromatography o (Forensics)
Compares fibers
found on a victim
- Used to analyze the
metal ions and organic o Detects pollution by
compounds in testing water
solutions. samples
- uses liquids that may
b) Liquid Chromatography incorporate
hydrophilic, insoluble
molecules
 - common type of o Separates amino
chromatography acids and anions
- paper is the stationary o RNA fingerprinting
c) Paper Chromatography phase o Separates and tests
- uses capillary action to histamines and
pull the solutes up antibiotics
through the paper and
separate the solutes
o Detects pesticide or
- simple and quick insecticide residues
d) Thin-layer method to check the in food
Chromatography purity of the organic o Analyzes the dye
compounds composition of fibers
(forensics)

4) Distillation
This is the process by which organic compounds
7 (contain carbon) are purified.
This is advantageous in the sense that two different compounds have different
boiling points. Take, for example, two different liquids are present in a
homogeneous mixture (completely miscible, e.g., H 2O & alcohol). If these
compounds have different boiling points, one of the compounds will evaporate
first (volatile) before the other will.

5) Drying
This is defined as the process of vaporizing and removing water or other liquids
in a certain material to form a dry solid. Physiochemical transformations result
from the simultaneous heat and mass transfer (which are critical factors in the
drying process).

6) Grinding
This is the process of turning a material into small pieces from fine to coarse
using a certain device, i.e., Wiley Mill or a grinding wheel.

7) pH measurement
A pH is a measurement of how weak and strong a particular acidic or basic
substance. A substance with a pH of 7 is neutral, i.e., H2O. A pH lower than 7
means acidic, i.e., acetic acid found in vinegar. A pH higher than 7 is a base, i.e.,
sodium hypochlorite found in bleaching products.

In many studies, pH is an important parameter or variable. For example, pH is

an important parameter or factor that is measured to determine the condition of
water because of pollution. How do you relate pH measurement with pollution or
water quality? With the use of pH, one can measure how acidic or basic water is
to determine its water quality. Pure water has a pH of 7, a little lower or higher
than that can be considered normal. But when the pH goes significantly lower, it
means the substance is already acidic and basic when it goes far beyond pH 7. If
drinking water for example becomes acidic or basic only indicates that other
 components are visible. A pH of 7.4. of water in rivers means good quality. Any
higher or lower than pH 7 means harm to the organisms living in the water.

8) Weighing
This procedure shows how o use a balance to make a precise and accurate
measurement, i.e., mass. To do this, the use of measuring devices is very
essential.

Example (Measuring devices)

o Solids – triple balance, top-pan electric balance, analytical electric balance,

etc.
o Liquids or solids that do not holder together (i.e., powder, ice, etc.) –
receptacle, graduated cylinder, etc.

III. Activity Proper

Activity 2. Experiment @ the Lab

Objective: Identify the standard methods and techniques in performing
experiments.

Directions. Using the given statement/title in each number, try to identify the
laboratory techniques being described. Write your answer on a separate sheet of
paper.

1. A particular food color mixture was separated into two colors – red and blue.

2. The human blood needs to keep its pH approximately 7.4 to maintain stability to
avoid any complications from it.

3. Bioprospecting for Novel Halophilic and Halotolerant Sources of Hydrolytic

Enzymes in Brackish, Saline and Hypersaline Lakes of Romania

4. The tannins, theobromine, and caffeine are obtained from the tea leaves in
boiling water.

5. Alcoholic beverages like whiskey are produced from fermented grains like corn or
rye at distilleries.

6. In baking, a cake of different varieties in yellow, white, and pound cakes to

produce a cake of correct volume, grain, and texture, the amount of air into the
batter needs to be considered.

7. Natural and synthetic food coloring are used as additives to make food more
acceptable and popular. Non-permitted coloring agents are not added to the

10
 foods, instead, edible colors are. Thus, these coloring agents are identified and
quantified.

8. Pharmaceutical industries used UV and fluorescence spectrometers invariably to

provide detectors for the instrumental-based systems.

9. A baker wanted to know the amount of salt or sugar that might affect the color
of the cake that she will bake.

10. Removing water in wet lettuce with the use of a salad spinner

Questions
Directions: Answer the following on a separate sheet of paper.
1. Based on the activities that you have accomplished, why do you need to know
the different laboratory methods and techniques in performing experiments?

2. Among the laboratory methods and/or techniques you have learned, which
would you likely utilize in conducting experiments that would answer your
research questions?

IV. Reflection
Complete the statements below.

I understand_____________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
I don’t understand _____________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
I need more information about ___________________________________________________
_________________________________________________________________________________
_________________________________________________________________________________
V. Links and Other References
Experimental Methods. 2021. ScienceDirect ®. Retrieved on February 9, 2021, from
https://www.open.edu/openlearncreate/mod/oucontent/view.php?id=233&pri
ntable=1

Lahora, Oliver A. (2020). Scientific Research for Junior High School. pp. 45-46, 55,
137

Retrieved on February 9, 2021, from https://eda.nc3rs.org.uk/experimental-design-

unit

Retrieved on February 9, 2021, from https://www.pubmed.ncbi.nlm.nih.gov

Retrieved on February 9, 2021, from https://biotopics.co.uk/microbes.tech1.html

Retrieved on February 9, 2021, from https://www.microbiologyonline.org>file

Retrieved on February 9, 2021, from https://www.chemengonline.com/solids-

drying-basics-and-applications/?printmode=1

Retrieved on February 9, 2021, from https://www.google.com/amp/s/dictionary

.comcambridge.org/us/

Retrieved on February 9, 2021, from

https://www.sciencedirect.com/topics/agricultural-and-biological-
sciences/titration

Retrieved on February 9, 2021, from

https://chem.libretexts.org/Bookshelves/Organic_Chemistry/Book%3A_Organ
ic_Chemistry_Lab_Techniques_(Nichols)/

VI. Answer Key

Activity 1
1) Chromatography (Paper)
2) pH measurement
3) Microbiological technique
4) Extraction
5) Distillation
6) Aeration (Food)
7) Chromatography (Paper)
8) Spectrophotometry
9) Titration
10) Centrifugation

Questions
Students’ answers may vary.