Inverse PCR (or alternative) recommendations requested

martin LEACH leach at bu.edu
Thu Nov 30 15:44:57 EST 1995


Hi.

inverse pcr has given me the bulk of my sequence that I obtained for a
promoter region...

A kit from Clontech now exists using adaptor-ligated genomic DNA fragments...5
libraries made from different restriction enzymes...the ref they give for this
is

nucl acids res. Siebert p.d. et. al '95 23:6 (in press)

all this is in their april 1995 clontechniques....go to their web site and see
if it is there...

other techniques i have used include..

Panhandle PCR
RAGE (rapid amplification of genomic ends: mizobuchi et. al. biotechniques)
you can get the full ref for RAGE from http://www.biotechniques.com archive
but not the full article.

If you wish I can dig up the refs for all of them...but Inverse pcr worked the
best for me. 

Martin

Bernard Murray (bernard at elsie.nci.nih.gov) wrote:
: Hello all,
: 	I am considering using inverse PCR to attempt to locate transgenes
: in the mouse genome.  Before starting I thought I would pick your
: collective brains on the topic.  Is this the technique of choice or has
: it been superceded?  Can you use long range PCR to extend well into
: the flanks?  What's the longest fragment that has been amplified?.  Can
: conventionally purified genomic DNA be used or do special precautions
: have to be taken?
: 	I have read two book chapters and several original articles
: on the subject but was wondering how things work out at the practical
: level.  Any advice/warnings/pointers gratefully received.
: 		Bernard

: Bernard Murray, Ph.D.
: bernard at elsie.nci.nih.gov  (National Cancer Institute, NIH, Bethesda MD, USA)


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.....          Martin Leach                Email:leach at bu.edu
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