wrong orientation of ligated fragment

roger morton ttguy at bigpond.com
Mon Jun 7 07:37:49 EST 1999


Sean Yoder <syoder at chuma.usf.edu> wrote in message
<375A87AD.3E7DDAB2 at chuma.usf.edu>...
>  I doubt a eukaryotic promoter would
> allow transcription though...but I'm just learning this stuff so take ir
for what
> it's worth.

You would be wrong here. Some people (and I have done it once) screen for
antibiotic restance insertion into plant promoter cassettes by plating the
ecoli on double Ab selection.

Eg cloning a hyrgomycin coding region into a CaMV 35S cassette contained in
a pUC plasmid you can select for  the plasmids with inserts by selecting on
amp and hygromycin. It does not always work. Seems to depend on the exact
context of the clone.

Roger Morton





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