amplification of 1.8 fragment for cloning

David-Paul Minde via methods%40net.bio.net (by davidminde from gmail.com)
Sun Feb 7 14:27:37 EST 2010


if you want maximum fidelity (some two orders of magnitude better than
Taq), just use Phusion/Pfu ultra II fusion HS (and subsequently A-tail
with Taq or subclone in blunt vector) ... just works with good yields
in some 95% of cases- also with whole plasmids of some 15 kb .. Simply
increases fun with clonings.
cheers,
David


David Minde MSc (TUM)

Cellular Protein Chemistry Room 707
Department of Chemistry
 Faculty of Science
 Utrecht University
Krytgebouw
 Padualaan 8
 NL-3584 CH Utrecht
 The Netherlands

office phone +31 30 253 4105

Science is what happens while we are making other plans (~John Lennon)



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