Cloning PCR products
num208ww at MBDS.NRC.CA
Wed Jan 29 21:46:21 EST 1992
In article <9201280146.AA11363 at molly.sdsc.edu>, mws%molly at SDSC.EDU (Michael W. Smith) writes:
>I am designing a new set of primers for PCR analysis and planning to use
>restriction enzyme sites for very rare cutting enzymes. The products would
>then be forced into a vector at those two sites.
>Does anyone have tales of great success or woe with the ClaI or SacII??
>I am especially interested in knowing the number of extra bases
>incorporated into the 5' end of the PCR primer.
>Thanks for any help
Mike, I also have not tried ClaI or SacII but tried NdeI and XbaI
which had 8 bp tails at the 5' end. The NdeI would not cleave at all
until I actually cloned the fragment blunt ended then exised my Nde
Xba fragment. The Xba cleavage worked without incident.
Dr. Warren Wakarchuk Prefered email: wakarchu at biologysx.lan.nrc.ca
Alternate : NUM208WW at MBDS.NRC.CA
IBS-Protein Structure and Design
National Research Council Canada
Canada K1A 0R6 ***THESE ARE MY OPINIONS NOT THOSE OF NRC***
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