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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