Restriction assay on PCR products

scarr at kean.ucs.mun.ca scarr at kean.ucs.mun.ca
Tue Nov 2 07:58:35 EST 1993


Subject: Re: Restriction in a PCR tube

X-News: leif bionet.molbio.methds-reagnts:8046

From: kanyuka at uk.ac.afrc
Subject:Restriction in a PCR tube
>Date: Mon, 1 Nov 1993 13:43:00 GMT
Message-ID:<1993Nov1.144946.20346 at gserv1.dl.ac.uk>

>Hello netters!
>
>	Has somebody try to digest PCR product in PCR tube (I mean in PCR
>reaction mixture) with different restriction enzymes? 
>	

ort of. We took 15ul of the PCR reaction into a second tube, added 1 unit
of RE, let it digest overnight, ran the product out in agarose, and scored
it. We did this with two different REs and got reproducible patterns. 
Although my grad student did the work, I don't believe we even had to 
adjust the buffer salts (it worked the first time without adjustment).
Steve

*********************************************************************
Steven M. Carr
Dept. of Biology
Memorial University of Newfoundland
St. John's NF A1B 3X9
CANADA

(709) 737-4776 office / -4713 lab / -4000 FAX
scarr at kean.ucs.mun.ca
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