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Michel Paul

qPCR: do both primers have to contain a GC-clamp in order to be effective?

Angel Vasquez  Follow

You can have GC-clamps (3' terminal G/C) in both primers; this may overall improve the PCR efficiency. However you should take care not to put too many G/Cs. This may lead to stabilization of primer pairs, leading to formation of primer-dimers. Also see this post: When designing primers how important is the GC clamp?

GC-clamps are not always essential and you should look at other parameters too.

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Emmanuel Simiyu  Follow
So, in summary: A clamp in one primer is good for efficiency and clamps in both are better (with the right amount of G/C ofcourse). So I don't need a GC-clamp in both of them. Is this right?More
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Joyce Phelps  Follow
@TomKral Not strictly necessary but good if you have it in both.More
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