21.3 In vitro gene cloning - the polymerase chain reaction Flashcards
1
Q
what does in vitro mean?
A
outside living organism
2
Q
main points for PCR
A
- DNA double helix strand separated by heating
- cooled, primers annealed
- heated, free nucleotides joined using DNA polymerase
3
Q
what does PCR require?
A
- DNA fragments
- DNA polymerase
- DNA nucleotides
- thermocycler
- primers
4
Q
How do you separate the DNA double helix strands?
A
- Heat to 95
- H bonds broken between complementary base pairs
5
Q
How are the primers added?
A
- Cool to 55
- complementary base pair forms H bonds
6
Q
Why are the primers added?
A
allow the DNA polymerase to bind to the base sequence
7
Q
What are primers?
A
short sequences of nucleotides that have complementary base pairing to ends of sequences
8
Q
How are double helix strands reformed?
A
- heat to 72
- DNA polymerase extends strands using free nucleotides
9
Q
what DNA polymerase is used and why?
A
- Taq. DNA polymerase
- from an extremophile bacteria, survives high temperatures
