Nurturing Life Sciences
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#Unit 13. Methods in Biology
The PCR primers must not have self complementary regions as this would lead to hairpin formation within primer molecules and make then unsuitable for PCR.
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The partially hybridized primers may become extended by the DNA polymerase and are amplified; sometimes this may monopolize the PCR .
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The two primers used in a PCR reaction have identical Tm for successful and specific amplification.
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The efficiency of amplification declines with an increase in the length of the target sequence.
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Efficiency of PCR is affected by primer length; it declines if the primers used for PCR arc too long.
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The annealing temperature has a marked influence on PCR efficiency; a temperature higher than the ideal annealing temperature reduces PCR efficiency.
