International Scholarships

Wednesday, 10 October 2007

PCR and multiplex PCR guide

Generalities
Components, PCR cycle, vials

Annealing time and temperature

Choosing PCR primers
How to design the primers

Extension time and temperature

Reaction volumes
Do they influence the results?

DNA template concentration

Number of PCR products
Multiplexing. How ?

Taq polymerase(s)
Comparisons and concentration

Primer amount
How much primer?

dNTP concentration
Interaction with magnesium

PCR buffers
Comparisons and concentration

MgCl2 concentration
Interaction with dNTP and salt (PCR buffer)

Salt (KCl) concentration
Essential optimization factor

Gel electrophoresis
Agarose and polyacrylamide (PAA)

Designing PCR programs
Customize them for your purpose

Adjuvants in PCR
DMSO, glycerol and BSA

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