Sequencing and copying DNA - Interrupted PCR and electrophoresis

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  • Created by: Steff06
  • Created on: 18-06-16 13:08
What does the reaction mixture in PCR contain?
The enzyme DNA polymerase, copies of the single-stranded template DNA fragment, free DNA nucleotides and primers.
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What do some of the free nucleotides carry?
Some carry a fluorescent marker.
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What happens if these modified nucleotides are added to the growing chain?
The DNA polymerase is thrown off and no further nucleotides can be added.
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What is the step in interrupted PCR?
The primer joins (anneals) at the 3' end of the template strand, allowing DNA polymerase to attach.
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What does the DNA polymerase do?
DNA polymerase adds free nucleotides according to base-pairing rules so the strand grows.
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What happens if a modified nucleotide was added?
The polymerase enzyme is thrown off and the reaction stops on that template strand.
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What happens as the reaction proceeds?
Many molecules of DNA are made. Fragments generated vary in size.
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How do the fragments vary in size?
In some of them, the template strand has only 1 additional nucleotide added before the polymerase is added. In others, the template strand is completed.
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What happens to the final added nucleotide?
The final added nucleotide is tagged with a specific colour.
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What does a laser do as the strands run through the machine?
A laser reads the colour sequence from the strand with only a single nucleotide to the one with 2 nucleotides, then 3 then 4 e.t.c.
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What can then be displayed?
The sequence of colours and the sequence of bases.
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What was the aim of the Human Genome Project?
To determine the whole human genome sequence.
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Other cards in this set

Card 2

Front

What do some of the free nucleotides carry?

Back

Some carry a fluorescent marker.

Card 3

Front

What happens if these modified nucleotides are added to the growing chain?

Back

Preview of the front of card 3

Card 4

Front

What is the step in interrupted PCR?

Back

Preview of the front of card 4

Card 5

Front

What does the DNA polymerase do?

Back

Preview of the front of card 5
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