UNIT3- AOS1 REVISION EXAM QUESTIONS

DNA manipulation techniques and applications

 the use of enzymes to manipulate DNA, including polymerase to

8
synthesise DNA, ligase to join DNA and endonucleases to cut DNA

 the function of CRISPR-Cas9 in bacteria and the application of this

9
function in editing an organism’s genome

 amplification of DNA using polymerase chain reaction and the use of gel
10
electrophoresis in sorting DNA fragments, including the interpretation of
gel runs for DNA profiling

 the use of recombinant plasmids as vectors to transform bacterial cells as

11
demonstrated by the production of human insulin

VCAA 2002

Question 5
There were three suspects in an assault case. A forensic scientist found blood, other than the
victim’s, at the site. DNA was extracted from five blood samples.
• the victim
• the blood at the assault site (not the victim’s)
• the three suspects
Polymerase Chain Reaction (PCR) was used on the extracted DNA.
a. A DNA polymerase enzyme is involved in the PCR process. Explain the role of the polymerase
enzyme in PCR.

2 marks

One of the regions used in the forensic analysis was a short tandem repeat (STR) sequence of 4
bases, called HUMTHO1. This sequence, located on chromosome 11, has many alleles which
differ from each other by the number of times the sequence AATG is repeated. It was this region of
chromosome 11 which was amplified using PCR.
The amplified samples were loaded onto a gel and electrophoresis was performed to separate the
fragments of DNA.
b. Name two properties of the DNA fragments which allow them to be separated from each other
during gel electrophoresis.

2 marks

c. Why is there only one band in lane 2 but two bands in lanes 3, 4, 5 and 6?

1 mark
d. How many different alleles at the HUMTHO1 locus are represented on the gel in individuals
2, 3, 4, 5, 6?

1 mark
e. Which piece of DNA, A or B, has the greater number of the 4 base repeat sequence?

f. Which of the suspects appears to have committed the assault? Explain.

 VCAA 2005

Question 14
DNA was incubated with radioactive nucleotides. After one cycle of replication the distribution of radioactive
and non-radioactive nucleotides in the DNA would be
 VCAA 2006

Question 4
a. Describe the appearance of a bacterial plasmid.

1 mark
A bacterial plasmid was modified in the laboratory so that it contained a gene for an enzyme which provided
resistance to the antibiotic tetracycline.
Bacterial cells, which in their natural environment were sensitive to the antibiotic tetracycline, were mixed
with the modified plasmid. The bacterial cells were treated so that they could take up the plasmid.
b. What is the name of the process in which a bacterial cell takes up a plasmid and expresses the genes of
the plasmid?
With respect to the growth of bacteria the results of plates A and C are shown. On plate A there is a
continuous growth of bacteria over the surface of the agar. On plate C the colonies are distinguishable from
each other.
c. i. What result would you expect on plate B with respect to the growth of the bacteria?

ii. Explain your answer to c.i.

1+1 2 marks
d. Explain why there is a difference in the way the bacteria have grown on plates A and C.

2 marks
Total 6 marks
Question 5
'CC' for Carbon Copy is the name of the first cloned kitten born in 2001. The nucleus of a cat's egg cell was
removed. It was replaced by a nucleus from a somatic cell of a donor female cat. Once development
commenced the egg cell was transferred into a surrogate female.
a. What is meant by the term cloning?

1 mark
The diploid number of a cat is 38.
b. i. How many chromosomes would have been in the nucleus that was removed from the egg cell?

ii. Is CC male or female? Explain.

1+1 2 marks
To determine if CC was in fact a true clone, studies were made of specific variable regions in the DNA of the
donor, CC and surrogate.
The results are shown in the table.

Donor DNA CC DNA Surrogate DNA

DNA variable region
(size in base pairs) (size in base pairs) (size in base pairs)
1 164/164 164/164 166/166
2 222/222 222/222 218/218
3 196/198 196/198 194/200
4 154/160 154/160 160/162

c. For each region of DNA there are two values, for example, 164/164. Suggest a reason for this.

1 mark

d. In the case of DNA variable region 4 in the donor DNA, why are the pairs of values different? 1 mark
e. From the data it was concluded that CC was a true clone. Explain the evidence in the table that supports
this claim.

1 mark
Total 6 marks

VCAA 2007
 d. Draw an arrow on the right-hand side of the diagram to indicate the direction of movement of
the DNA fragments.
1 mark
e. What do the standards consist of, and what is their purpose?
Questi
on 18
β-
cells
in the
f. From these results, give a conclusion which could be drawn about the huma
sample taken from the crime scene. 2 marks n
pancr
eas
produ
ce
insuli
n.
mRN
A
g. What further action would you recommend to the forensic scientists transc
investigating this case? 1 mark ripts
from

VCAA 2008
these cells can be used to produce complementary (cDNA) copies of the active
gene.
The enzyme needed to produce cDNA from an mRNA transcript is
A. restriction endonuclease.
B. reverse transcriptase.
C. ligase.
D. RNA polymerase.
 In this procedure, scientists select a particular restriction enzyme from an available range.
a. Explain the reason for their choice.

Electrophoresis uses electrical current to sort DNA fragments.

b. i. Describe one characteristic of this sorting process. 1 mark

ii. Explain why the DNA of each individual produces a different pattern of fragments after gel
electrophoresis, even when the same restriction enzyme is used. 1 + 1 = 2 marks

Examine stages Y and Z.

c. Describe, at the molecular level, what is meant by the term ‘hybridised’. Why is it necessary to carry out
hybridisation?

2 marks
Total 5 marks
 VCAA 2009

Question 5
Some people prefer to eat Wagyu cattle because of the high level of marbling (fat) in the meat. Four separate
DNA markers are used to test for marbling in an animal. Tested cattle are scored on a scale of zero to eight,
eight indicating the highest degree of marbling.
a. What does the use of four markers suggest about the inheritance of this characteristic?

1 mark
A Wagyu breeder discovered a small number of individuals in her elite herd that were suffering from
Chediak- Higashi Syndrome (CHS). CHS is an autosomal recessive condition that can affect species other
than cattle. The breeder required further information.
Gene probing was used to target CHS1, the allele responsible for the condition. The genetic probes for the
Wagyu CHS locus were derived from human alleles.

b. Given that the gene probe for a human works for the Wagyu, what can you infer about the chemical code
for this allele?

1 mark
The Wagyu CHS1 allele was isolated and given a fluorescent tag. It was introduced into a yeast cell as a
large, independent, cytoplasmic chromosomal segment called a Yeast Artificial Chromosome (YAC). In
addition to the allelic DNA, a YAC includes a centromere and a replication sequence. The yeast cells are
then incubated in the presence of growth stimulants and given time to replicate.
This procedure is similar to genetic engineering of bacterial plasmids, however the YAC is able to contain
much larger pieces of DNA than a plasmid.

c. i. In the bacterial cell above, draw a plasmid in the blank box.

ii. Bacterial plasmids lack a centromere. Why are YACs made with a centromere?

iii. What term describes the process of copying a gene?

A test was developed to identify each of the normal and mutant alleles. Two cows were chosen for testing.
Cow X – a cow known to have the autosomal recessive disease CHS
Cow Y – a phenotypically normal cow with a family history of CHS
The CHS locus was isolated from each, amplified and then treated with FokI restriction enzyme which
recognises the nucleotide sequences
 1 mark

d. What mutation resulted in Patient X having breast cancer?

1 mark
A daughter of Patient X was also tested for the first segment of the allele.
e. Would you expect the result of her test to be green, red or black? Explain your
answer.

1 mark
Total 6 marks

VCAA 2010
Question 8
DNA includes sections that are called short tandem repeats (STR). Mutations in STRs occur, on
average, every 500 generations. Different numbers of these repeats have no obvious effect on the
individual.
a. What is the likely reason for this?

1 mark
A young man, Ben, wants to find out more about his genetic ancestry. He sends a sample of cells,
obtained from a swab of his mouth, to a laboratory. On receipt of the sample, the laboratory treats the
cells to release the DNA to enable identification of STR markers.
b. Name the process used to produce many copies of the STR markers.

1 mark
Each of the STR markers produced is labelled with a dye and subjected to gel electrophoresis. Five of
Ben’s STR markers were compared with three family groups who have the same surname as him. The
following gels resulted.

c. Explain which family is Ben’s most recent common ancestor.

 2 marks
Total 4 marks

VCAA 2012

Question 3
Genetic information in humans can be obtained by DNA profiling.
In DNA profiling, the polymerase chain reaction is used by a scientist to amplify a particular
sequence of
DNA.
a. Briefly describe the steps of this technique.
VCAA 2013
VCAA 2014
VCAA 2015
VCAA 2016
VCAA 2017 (Northern Hemisphere)
 VCAA 2017

Question 34
The process known as polymerase chain reaction (PCR) involves repeated
cycles made up of several steps. During PCR the
A. first step in each cycle is to anneal primers to the DNA at a low temperature.
B. temperature must be lowered to 37 °C before the beginning of each cycle.
C. second step in each cycle is to heat the DNA to a high temperature.
D. final step of each cycle involves the use of DNA polymerase.

Question 38
The diagram below represents a DNA molecule and the position of the recognition sites for
the restriction enzymes
BamHI, EcoRI, HaeIII and SalI.

Also shown is a diagram of an electrophoresis gel in which the lanes R, S, T and U show the
separation of DNA
segments resulting from digestion of the molecule with one of the restriction enzymes.
 Which of the following shows the correct match between the lane and the restriction enzyme
used to digest the DNA
molecule?

Which one of the following conclusions can be made using the information given?
A. Individual 4 is the father of Individual 5.
B. Individual 3 is the mother of Individual 4.
C. Individual 5 could be the child of Individual 1 and Individual 3.
D. The parents of Individual 4 could be Individual 1 and Individual 2.
Question 6 (5 marks)
DNA hybridisation is a molecular technique used to determine the evolutionary relationships
between species. DNA is extracted from each species and cut into fragments.
a. Outline the steps involved in the DNA hybridisation technique following the extraction
and cutting of DNA. 3 marks

b. How does the technique of DNA hybridisation enable scientists to determine the
evolutionary relationships between species?

Question 9 (7 marks)
Scientists use recombinant bacterial plasmids as vectors to transform bacteria for a range of
purposes in research and biotechnology.
a. What is meant by the term ‘vector’ in the context given? 1 mark

A particular bacterial plasmid contains recognition sites for the restriction

enzymes EcoRI, HindIII and BamHI, along with two antibiotic-resistant
genes, ampicillin resistance (amp) and tetracycline resistance (tcl), and an
origin of replication (ORI).
The diagram below shows the positions of these recognition sites and antibiotic-resistant
genes as well as the position of the origin of replication within this plasmid.

One purpose of using recombinant bacterial plasmids is to produce bacteria capable of

synthesising human protein.

b. The restriction enzyme BamHI was used to help insert a gene coding for a human
protein into this plasmid.
i. Describe how restriction enzymes such as BamHI are used to help insert a gene
coding for a human protein into this plasmid. 2 marks
 ii. Draw and label a diagram in the space below to show the position of the human
gene in this plasmid when BamHI is used. Include the position of the recognition sites
for the restriction enzymes EcoRI, HindIII and BamHI on the plasmid. 1 mark

c. After the scientists had carried out the steps required to make plasmids with the inserted
human gene, these plasmids were mixed with a culture of bacteria. This mixture was treated
so that these plasmids would move into the bacterial cells. Not all bacteria took up these
plasmids.
Explain how scientists use antibiotics to identify which of the bacterial cells have been
successfully 3
transformed with plasmids carrying the human gene. marks
VCAA 2018
Question 30
Which lane represents a sample that was loaded with DNA fragments of four different
lengths: 100 bp, 150 bp, 200 bp and 300 bp?
A. W
B. X
C. Y
D. Z

Question 31
Which lane contains the band that is closest to the negative electrode?
A. W
B. X
C. Y
D. Z
VCAA 2020
VCAA 2021
 ANSWERS

VCAA 2002

Question 5 a The polymerase enzyme catalyses the production of a new strand of DNA or is
0/2 47 involved in making multiple copies of DNA or amplification of DNA
1/2 38 and
2/2 15 DNA polymerase replicates the DNA by extending from the primer or by
(Average complementary base pairing or by using the original DNA as a template.
mark Some students incorrectly identified the enzyme and discussed the role of another
0.67) enzyme. Many other responses gave one part of the expected answer. Students need to
be reminded to use the number of marks allocated to the question as an indication of
the depth required in their answer.
b A DNA fragment will move according to its charge and molecular weight (size) or
0/2 37 DNA is negatively charged and moves to the positive pole; smaller DNA fragments
1/2 31 move further or faster than larger fragments.
2/2 32
(Average
mark
0.95)
c There is only one band in lane 2 because individual 2 is homozygous, the others on
0/1 70 the gel are heterozygous or the two fragments of DNA are the same size or the
1/1 30 number of repeats in the two fragments is the same.
(Average Students should be able to read results from a gel and make appropriate conclusions.
mark Many find this a difficult task, but this can be improved by attention to practical
0.30) experience in class.

d There are 5 different alleles at the HUMTHO1 locus represented on the gel.
0/1 63
1/1 37
(Average
mark
0.37)
e DNA piece A has the greater number of the 4 base repeat sequence.
0/1 23 The greater the molecular weight of the sample the smaller distance the sample will
1/1 77 move from the loading well.
(Average
mark
0.77)
 f The bands on the gel for suspect 5 match the sample of blood found on the victim,
0/1 62 which was not the victim’s blood (lane 3).
1/1 38 Students could not be given a mark for the correct identification of suspect 5. The
(Average mark was awarded for the explanation as to why suspect 5 appears to have committed
mark the assault.
0.38)

VCAA 2005

14 10 64 19 7
15 4 79 2 14
16 9 6 79 6
17 1 93 4 1
18 52 2 33 12

VCAA 2006

Questions 4a–b.
Marks 0 1 2 Average
% 36 54 10 0.8
Question 4a.
Circular

Question 4b.
Transformation

‘Heat shock’ was not an acceptable answer as this is a process of taking up the plasmid. Neither was
‘transfection’ as this applies to viral vectors.

Many students were aware that the required answer began with ‘trans’. Examples of common
incorrect answers included transduction, translocation and transgenic. Recombination was also a
common incorrect answer.

Question 4c.
Marks 0 1 2 Average
% 34 12 54 1.2
4ci.
No growth of bacteria.

4cii.
The bacteria are sensitive to the antibiotic tetracycline and therefore do not grow.

Question 4d.
Marks 0 1 2 Average
% 51 27 22 0.7
 • Plate A: The bacteria are able to grow as there is no tetracycline present.
• Plate C: Only those bacteria which take up the plasmid can grow in the presence of tetracycline.

It was important that students distinguished between Plates A and C, recognised the significance of the
presence/absence of the antibiotic and realised that only some of the bacteria were transformed. One mark was
awarded for each of the points above. Some students gave a specific percentage transformation which could
not be deduced from the information. Students need to be aware that if their answers include incorrect
information they may not receive full marks.

Question 5a.
Marks 0 1 Average
% 42 58 0.6
Making genetically identical copies (of organisms).

The key point was that the copies are genetically identical. Many students stated that the copies were
identical or phenotypically identical; these responses were not awarded a mark.

Question 5b.
Marks 0 1 2 Average
% 15 37 49 1.4

5bi.
19

5bii.
Female. The donor of the nucleus was from the somatic cell of a female donor.

A common incorrect answer was ‘The cell was an egg cell and this comes from females’.

Question 5c.
Marks 0 1 Average
% 85 15 0.2
The reason for the two numbers is that this locus is found on a pair of homologous chromosomes.

This part was not well answered. Many students incorrectly thought the two numbers were representative
of the two strands of DNA.

Question 5d.
Marks 0 1 Average
% 88 12 0.1
The different values indicate that the individual being tested is heterozygous for this locus, indicating that the
alleles are
of different lengths.

Most students attempted this question, however, many students again related the numbers incorrectly to the
two strands of DNA or made reference to the occurrence of mutations having occurred, which was also
incorrect.

Question 5e.
Marks 0 1 Average
% 26 74 0.8
CC is a true clone as the DNA of CC is identical to the donor DNA and not the surrogate.

This was generally well answered, however students who stated the DNA was almost identical could not be
awarded the mark.
 VCAA 2007

Questions 2
Question 2a.
Marks 0 1 Average
% 29 71 0.7
PCR (or Polymerase Chain Reaction)

Question 2b.
Marks 0 1 Average
% 48 52 0.5
Heating:
• to 90°C or greater
• to dissociate the strands
• to break the hydrogen bonds.

Heating had to be mentioned, and the effect it had on the process.

Question 2c.
Marks 0 1 2 Average
% 62 22 16 0.6

On the diagram or in the space below the question, students needed to show primers anneal, one per strand, at
opposite
ends (for the first mark), and show one of:
• primers attach at 3’ end
• temperature reduced to approximately 50°C
• DNA/Taq polymerase is added
• propagation occurs away from the primer.
Question 2d.
Marks 0 1 Average
% 30 70 0.7

The arrow needed to point towards the top of the page.

Students were required to know that DNA has a negative charge and hence would move towards the positive
electrode.

Question 2e.
Marks 0 1 2 Average
% 46 23 31 0.9
Both of:
• the standard consists of fragments of known length
• they are used to estimate the size of the samples.

To gain the two marks, a link had to be established between the comparison of known to unknown. Too
often students made vague statements such as ‘The standard is used to compare to the sample’.

Question 2f.
Marks 0 1 Average
% 61 39 0.4
A suitable conclusion could be:
• the sample is not from the victim
• the sample could be from either suspect.

To gain the mark here students had to make a valid conclusion rather than a simple observation such as ‘The
sample matched the suspects’. Some incorrect conclusions included ‘both suspects must be guilty’, ‘both
suspects are identical twins’ or even ‘the victim wasn’t at the crime scene’.

Question 2g.
Marks 0 1 Average
% 63 37 0.4
Any one of:
• apply the same process to a different gene locus
• use another suitable DNA technique, such as DNA sequencing
• use another forensic method, such as blood analysis or fingerprinting.

To gain the mark, a forensic, logical and feasible action had to be recommended. Incorrect suggestions
included ‘check alibis’ and ‘interview the victim’.

VCAA 2008
 18 3 57 9 31
19 14 62 20 4
When cutting occurs at point Q the first fragment is 3 kB
and the remainder is 7 kB. This larger fragment is then
20 3 4 37 55 cut at P, removing the smaller right hand fragment of 2
kB, therefore leaving a fragment of 5 kB, the size of the
segment between points Q and P.

Question 6
Question 6a.
Marks 0 1 Average
% 60 40 0.4
Restriction enzymes cut at a particular base sequence/recognition site.

Answers that referred to cutting at ‘a place or gene locus’ were not awarded a mark.

Many students incorrectly thought that specific restriction enzymes cut DNA into specific lengths.
Question 6b.
Marks 0 1 2 Average
% 29 42 30 1.1
6bi.
Either of:
• DNA is negatively charged and moves towards the positive electrode
• shorter DNA fragments travel further/more quickly through a gel.

6bii.
DNA of individuals is different, therefore will be cut at different places and produce fragments of different length.

Question 6c.
Marks 0 1 2 Average
% 61 27 12 0.5
Both of:
• the RNA probe is complementary to the single stranded DNA sample and these join together/hybridise
• this enables the DNA to be seen/identified.

Many students treated this question as ‘describe DNA hybridisation’ and in doing so, they ignored the instruction
to
‘examine stages Y and Z’.

VCAA 2009

Question 5a.
Marks 0 1 Average
% 58 42 0.4
The characteristic is controlled by many genes or is an example of polygenic inheritance.
This was not an example of multiple alleles.

Question 5b.
Marks 0 1 Average
% 57 43 0.5
The chemical code is identical/very similar.

Some students stated that the code was ‘similar’, which was not precise enough. Answers which made
inferences on evolutionary relationships were not awarded any marks as they were not addressing the
question. Some students added
‘due to the universal nature of DNA’; however, this was incorrect.

Question 5ci-iii.
Marks 0 1 2 3 Average
% 18 43 30 9 1.3

5ci.
A circle was to be drawn

5cii.
Either of:
• the centromere was required for spindles to attach
• to allow mitosis to occur.

5ciii.
Gene cloning

Gene replication and DNA replication were also accepted. PCR and transcription were incorrect answers and
replication was deemed too vague.

Question 5di-ii.
Marks 0 1 2 3 Average
% 39 25 16 20 1.2

5di.
Both of:
• heterozygous
• it has different alleles (sequences).

To gain the mark students had to state ‘heterozygous’ and identify the reason for their choice.

5dii.
Two bands needed to be illustrated:
• near but slightly above 100
• near but slightly above 50.

Some students included extra bands or only one band, indicating that the DNA had not been cut.

Question 5e.
Marks 0 1 Average
 % 52 48 0.5
The electrode had been connected the wrong way.

Question 7a
A single-stranded segment of DNA which is (radioactively) labelled

It was important that students’ answers distinguished a probe from a primer. It was clear that the concept of a
probe was not well understood.

Question 7b.
Marks 0 1 2 Average
% 31 43 27 1
Both of:
• green – indicates hybridisation with a normal allele
• red – indicates a specific mutation and hybridisation with that allele have

occurred. This was essentially a comprehension question.

Questions 7c.
Marks 0 1 Average
% 87 13 0.2
The joining of complementary DNA from different sources

Question 7d.
Marks 0 1 Average
% 75 25 0.3
G was replaced by A

A point mutation occurred. The specific type of mutation was a base substitution; however, in light of the
data, the best answer was: G was replaced by A.

Question 7e.
Marks 0 1 Average
% 80 20 0.2
Either of:
• green if she inherited a normal allele from her mother
• red if she inherited the defective allele from her mother. Students struggled to read and interpret this

question.
 VCAA 2010

3 10 4 4 82
Question 8a.
Marks 0 1 Average
% 70 30 0.3
The repeats occur in non-coding regions of DNA or mutations occur in introns.

A common incorrect answer was ‘the mutation is silent.’

Question 8b.
Marks 0 1 Average
% 31 69 0.7
PCR or Polymerase Chain Reaction

Question 8c.
Marks 0 1 2 Average
% 10 14 77 1.7
Family Y and one of:
 there is only one difference/mutation between Y and Ben
 Y has more or four out of five STR markers, the same as Ben
 X and Z have a greater number of differences than Y does to Ben.

VCAA 2012

Question 3a.
Marks 0 1 2 3 4 Average
% 30 11 14 23 23 2
 heat DNA to approximately 90 °C or to separate strands
 cool to approximately 50 °C or to anneal/attach primers
 heat to approx 72 °C or Taq/DNA polymerase copies strands
 repeat cycle.

Many students set out clear answers that demonstrated a sound understanding of the process. Many
other students missed out on full marks by not stating that the process needed to be repeated.
Question 3b.
Marks 0 1 2 3 Average
% 34 13 11 42 1.6

An outstanding sprinter would have only one band as only one allele is present/homozygous. A good
sprinter has two different alleles/heterozygous.

Students who could not correctly place bands on the gel were still able to receive two marks for a suitable
explanation.

2013
2014

2015

2016
 2017

VCAA 2017

34 7 13 15 65 0
35 5 62 12 21 1
36 18 78 2 1 0
37 23 4 2 71 0
38 11 59 20 9 1
39 12 3 17 68 1
Question 6
Part a. required a description of the process and part b. required a discussion of the results.
Many students answered part b. in part a. and then rewrote the answer in part b. It is
advisable that students read each part of the question before answering, to avoid wasting
valuable time.
Question 6a.

Marks 0 1 2 3 Average
% 34 18 31 17 1.3

All:
• DNA is heated to break the bonds/separate the strands.
• Single strands from both species are mixed together and cooled.
• The resulting hybrid DNA is then reheated and the temperature at which the DNA
becomes single-stranded is noted.
Many students described PCR (which could have scored one mark) or included a full
description in their answer of DNA hybridisation as outlined above (which could have scored
full marks);
however, this highlighted the need for students to read the question carefully and answer what
is
being asked and not include the irrelevant description of PCR. The answer space provided
should be used as an indication of the length of the required response.
Question 6b.

Marks 0 1 2 Average
% 54 29 18 0.7

The temperature at which the DNA becomes single stranded indicates the degree of
complementary base pairing and the higher the temperature the more closely related
the two species are.
Question 9a.

Marks 0 1 Average
% 79 22 0.2

A means of transferring/transporting foreign/desired DNA into a cell/organism

Question 9bi.

Marks 0 1 2 Average
% 36 27 38 1

The restriction enzymes are used to cut the ends of the plasmid and the desired gene so
that the gene will insert into the plasmid.

Marks 0 1 Average
% 44 56 0.6
Question 9c.

Marks 0 1 2 3 Average
% 44 21 24 12 1.1

Grow the bacteria on agar containing ampicillin.

Those bacteria that will grow on agar containing ampicillin have the plasmid with the human
gene included.
If plasmid is not taken up, those bacteria are killed by ampicillin.

VCAA 2018

28 6 6 1 88
29 64 10 2 23
30 1 1 9 1
7
31 21 75 2 2
 VCAA 2020

Question 11a.
Marks 0 1 2 3 4 5 Average
% 28 11 9 17 9 26 2.5

Any five of:

 Shorter fragments move further and longer fragments move lesser.

 Standard used as a comparison and reference.
 BamH1 cuts the DNA fragment in one place and produces two fragments.
 HindIII cuts the DNA fragment in two places and produces three fragments.
 Largest/smallest fragments are produced when cut by HindIII
 BamH1 fragment length are 4000bp and 5500bp and HindIII fragment length are 8000bp,1000bp
and 500bp.
 When the two endonucleases are present, four fragments are produced, sizes of 5500bp, 2500bp,
1000bp and 500bp are produced.

This question provided students with the opportunity to apply the information provided to the gel and then
analyse the results provided.

Question 11b.
Marks 0 1 2 Average
% 28 52 2 0.9
0
Any one of:

 Absence of bands for BamH1/Tube 1 may have been caused by incorrect loading of the DNA
sample into the well.
 Only one band for HindIII/Tube 2 may have been caused by restriction enzyme being denatured.

Question 11c.
Marks 0 1 2 Average
% 14 54 3 1.2
2
Two factors are:

 size or length of the fragments

 concentration or viscosity of the agarose
 voltage or power used.

The length of time was not a suitable response.

Question 11d.
Marks 0 1 2 Average
% 15 3 49 1.2
7
Some examples of suitable answers are:

 Making sure the electrophoresis equipment is turned off to avoid electrocution.

 Wearing safety glasses to protect the eye.
 Wearing gloves to avoid contamination by DNA or the chemicals being used.
 Correct disposal of materials.

This was well answered and demonstrated that students could apply their knowledge from practical work
in this situation.

VCAA 2021