Week 3 - PCR

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Last updated 10:03 AM on 8/6/26
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12 Terms

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Increase Ta

Increase specificity

Decrease yield

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Decrease Ta

Decrease specificity

Increase yield

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Too much Mg2+

Too stable

Incomplete denaturation

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Too low Mg2+

Dysfunctional DNA pol

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Agarose vs Acrylamide - Resolution

Low vs high

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Problems - Non Specific Amplification

Self priming

Low Ta

Too much Mg/primer/template

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Primer Length

Oligonucleotide

18-30 nt

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Why Low Ta Decreases Specificity

Primers stick to loosely to DNA - not perfect match

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Why High Ta Increase Specificity

Higher energy - primers must match perfectly

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PCR Process

94 - 5min

94 - 30

55 - 30

72 - 45

x 30

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Taq - Half Life

40 min

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Methylated Base in Prokaryotes

A