A person does a series of PCR reactions and comes to you for advice. She has pur

Question

A person does a series of PCR reactions and comes to you for advice. She has purchased three sets of primers, tried them all, but none of them worked. Examine each pair and explain why it did not work. She is using taq DNA polymerase, which does not proofread.

The template strand

Please note if the problem is in only one or both. My answers are 1.only b 2. both 3.only a

Could you please check if I made a mistake, give explanations. Also, if taq DNA polymerase could proofread, would anything change?

Primer a Primer b Pair #1 5' GTCCAGC 3' 5' CCTGAAC 3' Pair #2 5' GGACTTG 3' 5' GCTGGAC 3' Pair #3 5' GTCCAGG 3' 5' CAAGTCC 3'

Explanation / Answer

First of all, I suggest you that primer length should be 18-30 is good, and your primer length is quite less which is not fair. Anyway, I would suggest you to look at the Tm ( melting temperature of the primer) and do the gradient PCR. I hope it should work.

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