DNA > Biotech
Polymerase chain reaction - PCR
Kary
Mullis' Biography and Nobel Lecture
DNALC:
PCR
Animation
PCR OPTIMIZATION
COMPONENT
|
MIN |
STANDARD |
MAX |
| DNA |
<1 pg |
0.1 - 1 mcg |
>1 mcg |
Primers mcM
|
0.1 |
0.2 - 0.5 |
1 - 2 |
| Primers ng |
30 |
60 - 150 |
300 - 600 |
| Primers pm |
5 |
10 - 25 |
50 - 100 |
| dNTP each
mcM |
2-20 |
50 - 200 |
200 |
| Mg mM |
0.5 |
1.5 |
2.5 - 4 |
| Enzyme U/100 mcL |
1 |
2.5 |
5 |
Cycle N.
|
20-25 |
30-40 |
45-50 |
Agarose gel electrophoresis: (Marker)
PCR optimization.
Free article (Lorenz, 2012, with video):
Polymerase
Chain Reaction: Basic Protocol Plus Troubleshooting and
Optimization Strategies
Articles
about PCR in PubMed: Timeline.
RT-PCR IIb - RT-PCR MPL/NBL
MPL and B2M RT-PCR.
PCR Primer Design
We recommend reading the book by Paul Rabinow:
"Making
PCR: A Story of Biotechnology", University of Chicago Press,
1996.
PCR - Variations on the theme
- Taq
Polymerase activities
- Long PCR
- High-fidelity PCR
- Asymmetric PCR
- One-sided / Anchored PCR - T4
RNA Ligase
- Hot-Start PCR
- Reverse Transcription PCR (RT-PCR)
- Quantitative
PCR - Real-Time PCR
- Multiplex PCR
- Allele-specific PCR
- In-situ PCR
- Emulsion PCR - Digital
Droplet PCR - Fig.
3
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