inverse PCR

Malay curiouser at ccmb.ap.nic.in
Thu Apr 29 09:23:15 EST 1999


In this regard I have a question. Does the length of the DNA fragment has
anything to do with the efficiency of inverse PCR???

Malay

Malay Kumar Basu
Center for Cellular and Molecular Biology
Hyderabad 500007
I N D I A

Fax: (00-91)40-7171195
Phone: (00-91)40-7172241
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This was only a test. Had this been an actual tagline...
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curiouser at ccmb.ap.ni.in


-----Original Message-----
From: Dr. Duncan Clark <Duncan at nospam.demon.co.uk>
To: methods at hgmp.mrc.ac.uk <methods at hgmp.mrc.ac.uk>
Date: Thursday, April 29, 1999 3:03 PM
Subject: Re: inverse PCR


:In article <7g94re$tvt$1 at nnrp1.dejanews.com>, juamber at my-dejanews.com
:writes
:>Hi, just a possibility is that you did not perform your circularisation at
a
:>sufficiently low dna concentration. At high dna concentration, besides
:>self-circularised restricted fragments, you could also get fragments of
:>different sizes ligating together (which may also circularise afterwards).
If
:>this is the case, the fragments you are interested in (those containing
your
:>transposon) could be ligating to one or more other fragments from the pool
:>before circularising.
:
:This caught me out last week when I tried it for the first time.
:Repeating the ligation at a 100fold diln. gave much cleaner results.
:Still the odd RE digest out of 10 tried gives smears but the others gave
:clean single or double bands. This was on bacterial genomic. I would
:think it is a lot harder for human xsomal.
:
:Duncan
:--
:The problem with being on the cutting edge is that you occasionally get
:sliced from time to time....
:
:Duncan Clark
:DNAmp Ltd.
:Tel: +44(0)1252376288
:FAX: +44(0)8701640382
:http://www.dnamp.com
:http://www.genesys.demon.co.uk
:

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