The diagram shows a DNA profile taken from a cn8;4oj - m-rq vcljjk- lfpe9/rime scene.


Following are the DNA profiles of the suspects in this crime. Which one is the guilty individual?

  What is the function of the Polymerase Chain.o1re gz( jyk;avd 17h Reaction?

  Unlike the majority of other DNA polymerases, Taq polymerase, which iss(flaodp x4vest(b 6ti), (k h0dxs 4* used in PCR reactions can withstand high temperatures without denaturing. Whichpl(si,v fb64* d (sktxoh( d0x ta4)es of the following is a likely source for Taq polymerase?

  What bond is formed between twe-1ovaqk, *c qfmo7vwa)lnly/-jx 07o glucose molecules to form maltose?

Gel electrophoresis is a technique often used to analyse DNA mjwipv79+m3 .xkl+mhh eqdi5 f8r- tb(olecules. It includes a gel and an electric field. The diav m+8h-j i+3fb9wr md.57(qkixe ltp hgram below shows the outcome of gel electrophoresis.


1.DNA is a type of nucleic acid. State one location where DNA is found in cells.

2.Deduce, with a reason, which direction the DNA fragment moves through the gel.

3.Distinguish between the functions of DNA and RNA in unicellular organisms.

  Which event happens in the leading strand but not in m7j d4w*nom-nthe lagging strand?

  Which of these answers is true in relation to the polymerase chain .9;+ xuo4jaz. / cv,ip liojpareaction?

I. High temperatures are used
II. It produces exact copies of a segment of DNA
III. It requires DNA ligase and restriction endonucleases

Below is an image showi:c gxl f5 p,xvm:xhb66ng the results from a paternity test.

1.State the two properties of DNA fragments that allow them to be separated on a gel.

2.Outline the method used in DNA profiling in paternity testing to reach the stage shown in the diagram.

3.Deduce, with a reason, who is most likely to be the father of:

3.1.Child A

3.2.Child B

4.State one use other than paternity testing for DNA profiling.

5.Draw a circle around the heaviest DNA band in father 4’s sample.

6.Suggest why it is important to analyse the DNA of the mother, as well as the father in paternity testing.

7.State one potential source of error in DNA profiling.

  The diagram below shows the replication ofi+8plrks(n r.ld0lp jy uj 0s;, x1o5i DNA.


Which of the following are correct functions of structures P, Q and S?

  What distinguishes DNA polymerase from RNA polymmq+ ccp ;oe1,ierase?

  Which of the following are examples of discoveries made using radioactive izk* cpdu m6l)w0 cnzdy45 4:zhsotopes?)*dzu4m cdpylznc:60h 4zk5w

I. Discovery of the light-dependent photosynthesis pathways using the $^14$C and $^12$C isotopes.
II. Discovery of DNA as the main genetic material using the $^32$P and $^35$S isotopes.
III. Discovery of the semi-conservative replication of DNA using the $^15$N and $^14$N isotopes.

The diagram below shows a replicatit .i)sx -oxfb*on fork which forms during DNA replication.



1.On the diagram above, label the following structures.

1.1.The 5' and 3' ends of both template strands.

1.2.With the letter E, label the strand in which replication is continuous.

1.3.With the letter P, label the protein that keeps both strands separated.

2.Compare and contrast the role of DNA polymerase I and DNA primase.

  The table below shows the percentage of nitrogenous bases fosc +ygrr aeqv,3 yp,uij:5g:(und in the DNA of Escherichia coli bacteria.,qsriv3y:+p5e( ucrgj:g a,y
Adenine Cytosine Guanine Thymine
32% 18% 18% 32%
What conclusion can be made based on the data shown in the table above?

Data was collected on the protein composition of venom sam i-*mt3 dziwt6cfb na:p5:su9 y mc(:hples taken from the Malayan Krait (B. candidus) found in three localities; Malaysia, Indonesia and Thailand. All samples were carefully transported and preserved at teub(6s t:y-dc zp:3im*mnhitc5wf9: amperatures below 20$^{circ}$C. The graph shows the number of different types of protein found in each venom sample.


1.1.Identify the independent and dependent variables in this investigation.

1.2.Venom samples were collected from different snakes, in different localities. Suggest two controlled variables, other than temperature, that would support the collection of comparable samples.

Below are the results from gel electrophoresis of one group of venom proteins.


2.1.Identify the group of venom proteins shown.

2.2.Explain how proteins are separated using gel electrophoresis.

Different mice were envenomed with samples from each of the three localities. It was observed that the venom with the lowest neurotoxicity was from the Malayan krait sampled in Malaysia.

3.Suggest why it was important to transport venom samples at temperatures below 20$^o$C.2
4.List two advantages of using mice as model organisms for humans

The venom samples were further analysed, and it was suggested that the Three Finger Toxins group (3FTx) of proteins contributed significantly to the neurotoxicity of the venom. This prompted a closer analysis of the different types of 3FTx proteins found in each venom sample. The data was presented in a graph.


5.Based on the data, suggest which type of 3FTx protein is most likely responsible for differences in the levels of toxicity observed between the different venom samples. Justify your answer.

The diagram below shows the replicatio018 lnf rvdxd4n of DNA.

1.Describe the characteristics of the enzyme Taq polymerase.

2.Suggest what the consequences for a cell would be if transcription were to stop.

3.Outline the roles of all enzymes involved in the replication of DNA and in the synthesis of mRNA through transcription.

  Which row represents the steps of the2o:5oar h. 2 lux(yne2p))rf6hyzoa b polymerase chain reaction (PCR) in the corrzbeo 2f 5(n 2aap26)r :uh yyh.loxor)ect order?

1.Enzymes are needed for genetic modification. Outline the r-hna ,2)c ;e7wojep zeole of named enzymes in genetic modification.-h,)e; ez 2wpjeca7on

2.Outline the stages of the polymerase chain reaction (PCR).

3.Discuss the potential risks and benefits associated with Bt corn.

The image shows a basic principlea g13xtzo + 6spxik;z. behind the polymerase chain reaction (PCR). The use of PCRo x.t3gpx i1zks6 +;az revolutionized the biotechnology industry and opened up possibilities for many applications.

1.List three uses for the DNA copies that are produced.

2.Outline the function of DNA primers in the polymerase chain reaction.

3.Explain the reason for using Taq polymerase in the polymerase chain reaction.

4.Describe one application of the DNA profiles that are produced by PCR and visualized by gel electrophoresis.

The cloning of plasmids is an essential skill for many research proj jzfjfi4dd,x9 19ai* 1dbj8xw ects. There are various techniques9f8,dxbjw ia1fxj9idz* 4 1jd for this type of cloning, but the first steps involving the use of restriction endonucleases and DNA ligation are long and often inefficient. The development of more reliable and economical techniques has been important in increasing our ability to manipulate DNA.
In vivo techniques have been successful in yeast and E.coli using DNA fragments with overlapping nucleotide sequences. However, if the overlapping sequences are too long it becomes problematic and sometimes more expensive because of the size of the primers required. To try to increase the efficiency of this method, further research was carried out, using E. coli and DNA fragments with homologous ends prepared using the PCR technique from template plasmids using Q5 DNA polymerase. In the first experiment, 2.6 kB plasmids were formed inside E. coli from 2 DNA fragments with varying lengths of overlapping nucleotide sequences (nt OL). Successful recombinations could be identified, as only these plasmids led to the fluorescence of the cells.


1.Describe the pattern of successful recombinants for the E.coli colonies shown in the photographs.

2.What is the function of DNA polymerase in the PCR technique?

The cloning accuracy was calculated as the percentage of the total colony number that was fluorescent. The total number of colonies and the cloning accuracy for the different overlapping nucleotide lengths are shown in the table and graph below.


3.Calculate the number of colonies with successful DNA recombinations on the agar plate with 25 overlapping nucleotides.

4.1.Explain how DNA recombination occurs with overlapping nucleotides.

4.2.Suggest a reason for the results seen in the agar plates containing E.coli with DNA fragments with 0 or 6 overlapping nucleotides.

5.Suggest a possible reason for the high cloning accuracy with 9 overlapping nucleotides.

Cloning efficiency was then investigated with different numbers of DNA fragments which had 18 or 25 overlapping nucleotides. The results from the experiment are shown below.


6.From the photographs of the petri dishes, deduce what effect the number of DNA fragments has on the number of colonies with successful DNA recombinations.

7.Calculate the percentage increase in the number of colonies for the 25 nucleotide overlap compared to the 18 nucleotide overlap for the recombination of 2 DNA fragments.

The investigation was extended to see how plasmid size and the number of DNA fragments affected DNA recombination in E.coli. The graphs below show the colony numbers resulting from either 2 DNA fragments or 3 DNA fragments being joined together to form different-sized plasmids (measured in kilobases).



8.Compare and contrast the results for the number of colonies produced from 2 DNA fragments and 3 DNA fragments.

Lastly, the accuracy of the cloning of the plasmids in colonies was tested. Colonies containing the 3 different sized plasmids were selected at random, plasmids were extracted and these were tested using gel electrophoresis. The results of 6 plasmids from each of the 3 different sizes are shown below.


9.Identify the plasmid number and size that has not been cloned accurately.

10.Using all the results provided, discuss whether they support the hypothesis that the most efficient process for bioscience research using plasmid recombination and cloning in bacteria is to use 2 DNA fragments, with 25 nucleotide overlap to form 2.6 kB plasmids.

  A DNA profile was made using DNA from a crime scene. Locus TH01c0ceyl8,+ 4kgpd)qzw was used to identify whether a suspect’s DNA matched the crime scene DNA. The l)w,0yc8+4dk qez pgcsuspect’s two alleles of the gene at Locus TH01 are shown below.


The alleles TH01-6 and TH01-4 were separated and visualised with gel electrophoresis. The resulting gel, with two bands of DNA, is shown below.

What is the DNA in Band B and why is this DNA at this position?

  Which of the following is/are characteristic(s) of Taq polymerac9p5lxb0fat 1ae h;, use?
I. Requires a DNA primer
II. Works well at temperatures above 98°C
III. Used in gel electrophoresis to fluorescent tag DNA molecules

  The diagram below shows the DNA band pattern from the first two genebs.p p6 tr7gq9rations of a bacterial culture. Gene76 tsp .pbrg9qration numbers are indicated at the bottom of the diagram. Bacteria were grown before the experiment in a medium containing a heavy isotope of nitrogen, $^{15}$N, and then transferred to the medium with the light isotope, $^{14}$N which marked the beginning of the experiment (generation 0).

  The diagram shows a DNA profile of ln )sqv lm v:)8y:ave+a child and the profiles of three potential pairs of parents. Which pair(s) couln mvlve:8ly v:s)+aq)d be the actual parents of the child?