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Old 05-16-2014, 07:00 AM   #1
Neiltje
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Location: Belgium

Join Date: Jan 2014
Posts: 21
Default primer dimer problem in PCR using Illumina protocol

Hi!
I'm using the two step PCR Amplicon approach to to some targeted sequencing.
My PCR with my primers without adaptor sequences works just fine. But when I add adaptor sequences according to the protocol :
Append to 5 end of forward PCR primer:
5 TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-[locus specific sequence]
Append to 5 end of reverse PCR primers:
5 GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-[locus specific sequence]

all of a sudden, I only have primer dimers in my PCR product.
Anyone ran into this problem? How did you solve it?
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