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For a complete description of primers, PCR programs and a discussion of the PCR conditions please consult: Andrologia 26: 97-106 (1994) and Biotechniques 23: 504-511 (1997). Click here to get the Biotechniques paper in PDF format.


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PCR and multiplex PCR guide


Table below lists the parameters influencing the PCR reaction and indicates some PCR and multiplex PCR applications. Clicking on an item in the table, will take you to a detailed page, describing that particular issue.

Topics for PCR optimization

0. Standards: examples of multiplex reactions used in all experiments

1

Generalities
Components, PCR cycle, vials

9

Annealing time and temperature

2

Choosing PCR primers
How to design the primers

10

Extension time and temperature

3

Reaction volumes
Do they influence the results?

11

DNA template concentration

4

Number of PCR products
Multiplexing. How ?

12

Taq polymerase(s)
Comparisons and concentration

5

Primer amount
How much primer?

13

dNTP concentration
Interaction with magnesium

6

PCR buffers
Comparisons and concentration

14

MgCl2 concentration
Interaction with dNTP and salt (PCR buffer)

7

Salt (KCl) concentration
Essential optimization factor

15

Gel electrophoresis
Agarose and polyacrylamide (PAA)

8

Designing PCR programs
Customize them for your purpose

16

Adjuvants in PCR
DMSO, glycerol and BSA

Some applications of PCR and multiplex PCR

A

Microdeletion screening

D

PCR labeling

B

LOH

E

InterAlu-PCR

C

Quantitative multiplex PCR*

F

Degenerative priming PCR