PCR and multiplex PCR guideTable 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.
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Topics for PCR optimization |
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0.Standards: examples of multiplex reactions used in all experiments |
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1 |
GeneralitiesComponents, PCR cycle, vials |
9 |
Annealing time and temperature |
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2 |
Choosing PCR primersHow to design the primers |
10 |
Extension time and temperature |
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3 |
Reaction volumesDo they influence the results? |
11 |
DNA template concentration |
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4 |
Number of PCR productsMultiplexing. How ? |
12 |
Taq polymerase(s)Comparisons and concentration |
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5 |
Primer amountHow much primer? |
13 |
dNTP concentrationInteraction with magnesium |
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6 |
PCR buffersComparisons and concentration |
14 |
MgCl2 concentrationInteraction with dNTP and salt (PCR buffer) |
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7 |
Salt (KCl) concentrationEssential optimization factor |
15 |
Gel electrophoresisAgarose and polyacrylamide (PAA) |
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8 |
Designing PCR programsCustomize them for your purpose |
16 |
Adjuvants in PCRDMSO, glycerol and BSA |
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Some applications of PCR and multiplex PCR |
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A |
Microdeletion screening |
D |
PCR labeling |
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B |
LOH |
E |
InterAlu-PCR |
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C |
Quantitative multiplex PCR* |
F |
Degenerative priming PCR |
