blunt-ending PCR product

john hess jfhess at ucdavis.edu
Fri Mar 25 18:56:39 EST 1994


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> From: mcgraw at BSCR.UGA.EDU
> Newsgroups: bionet.molbio.methds-reagnts
> Subject: blunt-ending PCR product
> Date: 25 Mar 1994 14:07:59 -0800
> Organization: BIOSCI International Newsgroups for Molecular Biology
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> 
> 
> Hi,
> 
> does anyone out there care to recommend or reference a sure-fire method
for
> making a PCR product ready for ligation with a linker via blunt-end
ligation?
> In other work, we've used primers with restriction sites and we've done
T-A
> cloning also...this is something different...we really need to ligate
linkers
> to PCR products via blunt-end ligation.  Any suggestions?
> 
> Thanks,
> 	Al McGraw



I don't work for pharmacia but can say I've had good luck with the
sureclone kits they sell.  Not cheap, $225 for 25 rxns but once your sure
your blunt ended CIAP treated vector doesn't self ligate, don't do a
vector alone ligation control. 

If someone else can post another sure fire method, I'd love to try it and
switch to save some bucks but for now, I clone all my PCR products into
Sma cut pSP72 with sureclone. 

John Hess, PhD                    Phone me 916 752 8420
Dept of Human Anatomy             FAX me 916 752 8520
University of Calif               Email me jfhess at ucdavis.edu
Davis, CA                         or leave me alone, your choice.



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