PCR Mutagenesis

Bernard Lassegue medbpl at emory.edu
Fri Jul 21 18:51:42 EST 1995


In article <3u6pii$ovp at cisunix1.dfci.harvard.edu> Shoumo Bhattacharya,
sbhattac at mbcrr.harvard.edu writes:
>You need to have convenient restriction sites around the site so
>that the pcr products are about 3-500nt. I use Pfu polymerase, and very
>low concs. (20-50uM) dntp; this reduces artefacts. You need about 12-15
>nt on either side of the mismatched site, so that the pcr can be done
>at relatively high temperatures, again this reduces artefacts. Use
>plenty of plasmid as template; this reduces the number of cycles you
>need.  The actual stuff can be done in a day.

Could you indicate what is your PCR mutagenesis method and recommend a
bibliographic reference?
Thanks
Bernard
medbpl at emory.edu



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