PCR Revision

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 RNA must be converted to DNA because PCR requires the use of taq
polymerase which is a DNA polymerase

The purpose of DNA amplification in PCR is to create more DNA samples, allowing
more tests to be run

Each stage requires a different temperature due to the purpose. At the

denaturation stage, the hydrogen bonds between DNA must be broken. As
hydrogen bonds are relatively strong intermolecular forces, this stage requires a
high temperature of 95 degrees. The next stage of annealing involves the
attachment of primers, therefore the temperature needs to be lowered to 50-60
degrees. The final stage of extension involves the attachment of free nucleotides,
which is catalysed by taq polymerase, therefore requires the optimum
temperature of 72 degrees

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Denaturation: Hydrogen bonds between two DNA strands are broken at

95 degrees

Annealing: Attachment of primers at 50-60 degrees

Extension: Attachment of free nucleotides catalysed by taq polymerase

at 72 degrees

1. Charge . DNA is negatively charged therefore is attracted to the cathode

2. Size. Smaller DNA fragments move faster through the agarose gel

3. Amount of time voltage is applied for

4. Concentration of agarose gel (denser agarose results in the molecules

moving more slowly)
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