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Old 12-20-2018, 08:50 PM   #1
Kujin Kwon
Junior Member
 
Location: Korea, South

Join Date: Dec 2018
Posts: 3
Default PCR after size selection of cDNA library

Hi all,

Recently I'm trying to make cDNA library using Tag-seq protocol.
After amplifying my sample with illumina adapter, I select 300~500bp size (0.5x TAE, Zymo Gel DNA recovery kit)
However, I found that sample concentration is too low, so I re-amplified them with illumina adapter but, concentration was lower than before (PCR doesn't work)

-> Size select before adding adapter is impossible (too low concentration)
-> Gel elution kit work well with other experiment (cloning, etc...)
-> My PCR condition : (98C 30sec, [98C 10sec, 72C 30sec, 72C 60sec] , 72C 5min) This condition works well in first amplification.

Any comments would be welcomed.
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