21.3 In vitro gene cloning - the polymerase chain reaction Flashcards

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1
Q

what does in vitro mean?

A

outside living organism

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2
Q

main points for PCR

A
  • DNA double helix strand separated by heating
  • cooled, primers annealed
  • heated, free nucleotides joined using DNA polymerase
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3
Q

what does PCR require?

A
  • DNA fragments
  • DNA polymerase
  • DNA nucleotides
  • thermocycler
  • primers
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4
Q

How do you separate the DNA double helix strands?

A
  • Heat to 95
  • H bonds broken between complementary base pairs
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5
Q

How are the primers added?

A
  • Cool to 55
  • complementary base pair forms H bonds
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6
Q

Why are the primers added?

A

allow the DNA polymerase to bind to the base sequence

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7
Q

What are primers?

A

short sequences of nucleotides that have complementary base pairing to ends of sequences

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8
Q

How are double helix strands reformed?

A
  • heat to 72
  • DNA polymerase extends strands using free nucleotides
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9
Q

what DNA polymerase is used and why?

A
  • Taq. DNA polymerase
  • from an extremophile bacteria, survives high temperatures
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