PCR and multiplex PCR guide
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 |
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