PCR Cont. Flashcards
1
Q
Equation for PCR Theory
A
Nf=N0(1+Y)^n
2
Q
Y is the efficiency of the _____. It is a decimal from why?
A
amplification per cycle. Decimal from 0-1 not a perfect 2^n
3
Q
What is Taq polymerase?
A
A thermostable polymerase
4
Q
How is thermal stability achieved in taq polymerase?
A
- Hydrophobicity of the core of the enzyme
- Stabilization of electrostatic forces
5
Q
Why is taq pol important?
A
It stays active and stable with heating and cooling (2-4)
6
Q
What is the optimum temperature of taq and common temp?
A
75-80 but common is 68-72
7
Q
What does taq need?
A
ssDNA, primer with 3’OH
8
Q
What type of exonuclease activity does taq have?
A
5–>3 but not 3–> so cannot correct mistakes
9
Q
What does the 5-3 exonucelase activity allow taq to do?
A
It allows it to remove ahead of the growing template
10
Q
What does a primer determine?
A
specifity of amplification
