New Senior Secondary Mastering Biology (Second Edition)

Practical Worksheet for SBA E4

Suggested answers to Practical Worksheet

Ch 1 Techniques in modern biotechnology

Practical 1.1 Using polymerase chain reaction to amplify DNA

fragments
Understanding procedure (p. 104)
1 Free nucleotides act as the building blocks of the newly synthesized DNA strands.

2 Heat-stable DNA polymerase is not easily denatured by heat. It can catalyse the synthesis
of new DNA strands at high temperature.

3 To collect all the reaction mixture from the inner wall of the PCR tubes.

4 To avoid degradation of the PCR products at room temperature.

Results (p. 105)

1 (Results vary with Ss.)

2
PCR tube DNA band

A +

B +

C -

Discussion (p. 106)

1 For PCR tube A, there are DNA bands on the gel because the DNA template is amplified.
The primers anneal to the DNA template and new DNA strands are synthesized from free
nucleotides under the action of the DNA polymerase.
For PCR tube B, there are DNA bands on the gel because the control DNA is amplified.
The primers anneal to the control DNA and new DNA strands are synthesized from free
nucleotides under the action of the DNA polymerase.
For PCR tube C, there are no DNA bands on the gel. This is because there is no DNA
template for the annealing of the primers. Therefore, no amplification occurs.

 Oxford University Press 2015

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 New Senior Secondary Mastering Biology (Second Edition)
Practical Worksheet for SBA E4

2 Inaccurate volumes of reagents are transferred due to pipetting errors. / Some reagents
remain on the inner wall of the PCR tubes before PCR.

3 Prewet the micropipette tip with the reagent before use. / Centrifuge the reaction mixture
for a longer period of time to collect all the reagents from the inner wall of the PCR
tubes before PCR.

Extended question (p. 106)

There would be no DNA bands on the gel. This is because the two DNA strands of the DNA
template cannot separate below 72 °C. Therefore, amplification would not occur.

 Oxford University Press 2015

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