Help in Primer designing for expression...

[Duncan Clark]

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 
literary masterpiece:
>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.

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

Duncan Clark
GeneSys Ltd.