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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