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mutagenesis

mutagenesis - Protocols and Methods Forum

mutagenesis - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 08-12-2008, 12:31 PM
David-Paul Minde
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Default mutagenesis



.... (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 ...

best,

David

--
David Minde MSc (TUM)

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

office phone +31 30 253 4105

private address:
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3572 TW Utrecht

I never think of the future. It comes soon enough.
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Res severa verum gaudium. (~true delight is a severe issue)
Seneca
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  #2  
Old 08-12-2008, 08:18 PM
DK
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Default mutagenesis

In article <[Only registered users see links. ].net> , "David-Paul Minde" <[Only registered users see links. ]> wrote:

Actually, I believe the highest fidelity is PfuUltra Fusion.
Basically, Stratagene stole Finnzyme's idea, somehow
managed to circumvent IP (patent) protection and ended
up making a better product.

DK
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  #3  
Old 08-14-2008, 08:27 AM
Duncan Clark
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Default mutagenesis

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 :-)

Duncan

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I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.
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  #4  
Old 08-14-2008, 02:43 PM
DK
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Default mutagenesis

In article <GZncfDEjx+[Only registered users see links. ].com>, Duncan Clark <[Only registered users see links. ].uk> wrote:

Thank, Duncan! Interesting info.

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