IN VITRO CLONING Flashcards

1
Q

In vitro amplification

A

in vitro amplifcation uses polymerase chain reaction.
dna fragments can be amplified using in vitro cloning.

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2
Q

in vitro cloning

A

copies of dna are made outside the living organisms using polymerase chain reaction.

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3
Q

Stages of pcr

A
  1. a reaction mixture is set up that contains a dna sample, free nucleotides, primer and dna polymerase ( TAQ polymerase)
  2. the mixture is then heated to 95 degree celsius to break the hydrogen bonds between two dna strands.
  3. the mixture is cooled down to 50 - 65 degree celsius so that the primers can bind to the strands.
  4. reaction mixture is heated again to 72 degree celsius so dna polymerase can work.
  5. dna polymerase lines up free dna nucleotides alogside each template strand with complementary base pairing
  6. two new copies of fragments of dna are formed and one pcr cycle is complete.
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4
Q

advantages of pcr

A
  1. automated so more efficient
  2. 100 billion copies can be made in a an hour
  3. doesnt require living cells - quicker and less complex techniques are needed.
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5
Q
A
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