Inverse PCR (or alternative) recommendations requested
leach at bu.edu
Thu Nov 30 15:44:57 EST 1995
inverse pcr has given me the bulk of my sequence that I obtained for a
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
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..
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.
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: it been superceded?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 Murray, Ph.D.
: bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
..... Martin Leach Email:leach at bu.edu
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