Reamplification problem of the eluted PCR amplicons from PAGE gel
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yphong at YUMC.YONSEI.AC.KR
Fri Dec 6 21:08:13 EST 1996
I am working on DDRT-PCR with amoebas. Currently, I found bunch of PCR
products which showed differential display between compared mRNA
populations. I eluted target amplicons from dried 6% polyacrylamide gel
containing 7M urea according to Maniatis' protocol and tried to reamplify
target DNA. Here, I am in trouble of reamplifying the target DNA of the same
size in the original PAGE gel. I had modified the PCR conditions but I could
not get the exact target amplicons. Sometimes I got smaller amplicons than
the target, which were not present in the negative control reactions with no
DNA. Please, help me to solve this problem.
Thanks in advance.
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