Reamplification problem of the eluted PCR amplicons from PAGE gel

»ýÃæÇб³½Ç ±³¼ö È«¿ëÇ 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.
YPH 



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