[Bioforum] normalized cDNA
(by ilange from wsu.edu)
Tue Sep 25 16:37:41 EST 2007
I recently purchased a kit from Evrogen (Trimmer) to create a normalized
cDNA library. I made two attempts. The first time I followed their
general instructions closely and ended up with nothing but repetitive
concatamers (basically resembling my PCR primers) after sequencing.
The second approach was done very slowly, I monitored every step and
checked for RNA quality, mRNA quality and cloned the resulting cDNA into
a vector for sequencing before using the kit. Everything was fine until
I used Trimmer. I ended up with the same nothing!
I talked to somebody who tried it himself and had after 6 month of labor
and costs nothing. He recommended to send our DNA to a service lab. But
we do not have the funds and need the normalized cDNA.
Is there anybody who had with some success with this kit? Some advice
how to make it work?
Thanks for your help!
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