Would exonic primers affect gDNA amplification?

  • Thread starter Wek
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Wek

18
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Will a primer designed for an exonic sequence anneal and sucessfully amplify a gDNA strand? I'm having trouble amplifying gDNA using that primer. cDNA with that primer amplifies perfectly. I have no problem amplifying gDNA using another primer that anneals to intronic sequences.
 
44
0
I can't think of a reason why any given primer should work only for cDNA. Maybe the reason why you're having trouble with the genomic DNA amplification has to do with other possible annealing sites on the genomic DNA? I had this problem before. If you have upstream/downstream genetic information, maybe you should try a nested PCR?
 

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