SmaI site gone!?
BMBTJY at leeds.ac.uk
Mon Sep 22 08:45:34 EST 1997
A strange thing has happened. I subcloned a cDNA sequence into a eukaryote
expression vector using flanking SmaI restriction sites. I've used this
construct for expression and everything seems to be working fine. However,
I've done some SDM on the gene in this vector and I now want to subclone it
into a different plasmid. Well, seeing as I ligated it in at SmaI sites I
thought I would excise it at the SmaI sites but they appear to have gone!?
Now the only thing I can think of is that I had some exonuclease activty
arround when I did the ligation which has chewed back the SmaI recognition
site. Is this something other people have seen? Or could there be another
explanation? Thanks in advance.
Tim Young #8-)
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