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.
2
Q
in vitro cloning
A
copies of dna are made outside the living organisms using polymerase chain reaction.
3
Q
Stages of pcr
A
- a reaction mixture is set up that contains a dna sample, free nucleotides, primer and dna polymerase ( TAQ polymerase)
- the mixture is then heated to 95 degree celsius to break the hydrogen bonds between two dna strands.
- the mixture is cooled down to 50 - 65 degree celsius so that the primers can bind to the strands.
- reaction mixture is heated again to 72 degree celsius so dna polymerase can work.
- dna polymerase lines up free dna nucleotides alogside each template strand with complementary base pairing
- two new copies of fragments of dna are formed and one pcr cycle is complete.
4
Q
advantages of pcr
A
- automated so more efficient
- 100 billion copies can be made in a an hour
- doesnt require living cells - quicker and less complex techniques are needed.
5
Q
A