True whole-genome amplification

phd at mrc-lmb.cam.ac.uk phd at mrc-lmb.cam.ac.uk
Fri Nov 28 12:29:22 EST 1997


My group has a long-standing interest in developing a true whole genome
PCR amplification. The requirement is to amplify, from a single
mammalian genome's worth of DNA (<3pg), all sequences by >10E3-fold.  No
existing WGPCR acheives this.  

We have already tried: Telenius PCR (good amplification of <20% of
genomic sequences; no amplification of others); N15-primed PCR
(amplification of all sequences, but only by 100-1000 fold; no ability
to re-amplify further); Bohlander primer (similar results to Telenius);
PEP (same problem) and others.  Our best method so far gives us a
capture of about 50% of sequences (initially present as single
molecules) with >10E4-fold amplification.

If anyone is interested in similar problems and would like to discuss
this further, please get in touch.



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