T-cloning vs. Srf-cloning?
kvernick at helix.nih.gov
kvernick at helix.nih.gov
Sun Dec 11 21:21:12 EST 1994
Netheads,
Does anyone have an idea of the relative efficiency of cloning PCR products by ligation
into mono dT-tailed vectors compared to ligation of blunt molecules in the presence of
blunt-cutting restriction enzyme (a la the SrfI system of Stratagene)?
Both approaches work fine for obtaining clones containing a predominant PCR fragment, but
this is a pretty non-demanding application. I need to generate clones from a mixture of
PCR products, so the important parameters are the number of clones generated per unit
mass of PCr products, and the proportion of colonies harboring non-recombinant plasmids.
Any ideas?
thanks,
Ken Vernick
NIH
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