directional cDNA cloning
K J C 944
kjc944 at aol.com
Sun Jan 7 00:54:42 EST 1996
Mayumi,
I have also used Stratagene's cDNA synthesis kit with their Zap Express
vector with great success. I screened with an antibody and found several
Hundred (yes, hundred) positive clones per plate. One thing I tried that
may be of interest to you is that instead of using P32 labelled nt, I
stained my gels with SYBR Green (Molecular Probes) to visualize both first
and second strand synthesis. To do this, I had to double the
concentration of mRNA and used double the volume of reagents so that I
would have enough product. This works great as long as you have enough
starting material.
(No relation to Stratagene or Molecular Probes, Just a satisfied
customer.)
Good luck, Kathy
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