PCR Flashcards
1
Q
PCR components
A
- DNA primers - 3uM
- Taq DNA polymerase
- dNTPs
- buffer 10x
- nuclease free H2O
- template DNA
- MgCl2
2
Q
PCR process
A
- Denaturation - 95C - 30 seconds
- Annealing - depends on primer melt (within 2-3C) - 30 seconds
- Elongation - 72C - 1 kb/min
3
Q
DNA primers
A
15-40 bases
4
Q
Taq DNA polymerase
A
builds the DNA at high temp
5
Q
dNTPs
A
building blocks of DNA
6
Q
buffer
A
suitable environment for Taq polymerase
7
Q
nuclease free H2O
A
to mix with conc. buffer
8
Q
template DNA
A
original DNA
9
Q
MgCl2
A
works as a cofactor for Taq polymerase to make it work
10
Q
amplification formula
A
Nt = No X 2^n Nt = # DNA after cycles No = # DNA before cycles n = # of cycles
11
Q
control group
A
has no template DNA to make sure there is no contamination in the sample
12
Q
Formula for measurements
A
M1V1 = M2V2
