Help in Primer designing for expression...
(by blackhole from abuse.plus.com)
Mon Jul 20 08:22:01 EST 2009
Historians believe that in newspost
<mailman.618.1248049891.21502.methods from net.bio.net> on Sun, 19 Jul 2009,
Dr. Hiranya S. Roychowdhury <hroychow from nmsu.edu> penned the following
>Since the ATG start codon is part of the NCO1 sequence, we use that RE
>site for in-frame ligation. So, if your amplicon is digested and
>religated at the NCO1 site, you should be fine.
Just remember that for Nco I, the codon following the ATG must start
with a G either original or you must add one i.e. CC - ATG - GNN
Also pick the correct pET21 i.e. pET21d :-)
For this reason I much prefer to use Nde I whenever possible i.e.
CATATG. No problems with extra aa's.
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
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