U3A1: 4C polymerase chain reaction (PCR) Flashcards

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1
Q

how does PCR begin

A

a mixture of a DNA sample, taq polymerase, nucleotide bases and DNA primers are placed into a thermal cycler.

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2
Q

purpose of PCR

A

to amplify a sample of DNA by creating additional copies via thermal cycling.

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3
Q

explain first stage of PCR

A
  1. denarutation:
    DNA is heated to 90-95 degrees to break the hydrogen bonds, making single-strand DNA.
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4
Q

explain second stage of PCR

A
  1. annealing:
    single-stranded DNA is cooled to 50-55 degrees to allow primers to bind to complementary sequences.
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5
Q

explain third stage of PCR

A
  1. elongation:
    DNA is heated to 72 degrees so taq polymerase can work optimally, binding to the primer, acting as a starting point for the synthesis of the new complementary strand of DNA.
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6
Q

forward primers

A

binds to the start codon at the 3’ end of the template strand.
causes taq polymerase to synthesise a new strand of DNA in the same direction RNA polymerase would function.

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7
Q

reverse primers

A

binds to the stop codon at the 3’ end of the coding strand.
causes taq polymerase to synthesise a new strand in the reverse direction that RNA would function.

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