Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum

Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum (http://www.molecularstation.com/forum/)
-   Protocols and Methods Forum (http://www.molecularstation.com/forum/protocols-methods-forum/)
-   -   mutagenesis (http://www.molecularstation.com/forum/protocols-methods-forum/20029-mutagenesis.html)

David-Paul Minde 08-12-2008 12:31 PM

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
Krytgebouw
Padualaan 8
NL-3584 CH Utrecht
The Netherlands

office phone +31 30 253 4105

private address:
Griftkade 4 bis
3572 TW Utrecht

I never think of the future. It comes soon enough.
Albert Einstein
Res severa verum gaudium. (~true delight is a severe issue)
Seneca

DK 08-12-2008 08:18 PM

mutagenesis
 
In article <[Only registered and activated users can see links. Click Here To Register...].net> , "David-Paul Minde" <[Only registered and activated users can see links. Click Here To Register...]> 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

Duncan Clark 08-14-2008 08:27 AM

mutagenesis
 
Historians believe that in newspost <Remok.20246$[Only registered and activated users can see links. Click Here To Register...]>
on Tue, 12 Aug 2008, DK <[Only registered and activated users can see links. Click Here To Register...]> 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

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

Duncan Clark
GeneSys Ltd.

DK 08-14-2008 02:43 PM

mutagenesis
 
In article <GZncfDEjx+[Only registered and activated users can see links. Click Here To Register...].com>, Duncan Clark <[Only registered and activated users can see links. Click Here To Register...].uk> wrote:

Thank, Duncan! Interesting info.



All times are GMT. The time now is 07:55 PM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved

Page generated in 0.09024 seconds with 11 queries