How to cut PCR product with restriction enzyme ?

Greg Gloor ggloor at julian.uwo.ca
Fri May 27 12:20:52 EST 1994


How to cut PCR product with restriction enzyme ?

I do a PCR reaction in 15 ul. You must optimize the reaction to give a good
yield. I then add the minimum volume of a solution that contains 2-3 units of
enzyme and that brings the MgCl2 and NaCl concentration to that needed by the
restriction enzyme. Digest for 2 hours and run on the gel. I almost never have
partial
digests. This has worked fine for HaeIII, NdeII, RsaI, EcoRI, HindIII, PstI
BamHI.
Regards..........Greg Gloor           ---------------------->       ---------
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voice 519-661-3526  fax 519-661-3526  ---------     <------------------------



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