.... (hope you have no longer any problems with mutagenesis as in a post some
6 years back but ..) I strongly prefer Phusion (=highest fidelity
commercially available polymerase) PCR for mutagenesis. Piece of cake easy
for up to ~20 kb plasmids ...
David Minde MSc (TUM)
Cellular Protein Chemistry Room 707
Department of Chemistry
Faculty of Science
NL-3584 CH Utrecht
office phone +31 30 253 4105
Griftkade 4 bis
3572 TW Utrecht
I never think of the future. It comes soon enough.
Res severa verum gaudium. (~true delight is a severe issue)
Historians believe that in newspost <Remok.20246$[Only registered users see links. ]>
on Tue, 12 Aug 2008, DK <[Only registered users see links. ]> penned the
following literary masterpiece:
MJ Bioworks (prior to MJ's downfall over the thermal cycler infringement
with ABI) patented Sac 7/Sso7 (7kd non-specific ds DNA binding protein
from Sulfolobus) as a fusion with a DNA polymerase. They then specified
in another patent that one had to have various aa's at positions a ... z
in the Sso sequence in order to bind DNA.
Stratagene basically altered every other amino acid else bar those and
after lots of screening identified a modified Sso7 that worked as well
as the wt Sso7 in a fusion.
All fun and games :-)
I love deadlines. I especially like the whooshing noise they make as
they go flying by.