Dear Netters:
I am screening A. thaliana genomic library with a heterologous probe. By
screening about
160'000 plaques(!), I have got 9 independent clones. These clones are purified
to homogeneity by 3-5
time screening. All washes were to high stringency. To double check that these
(or at least one of them)
are corresponding to the right gene and not other related genes (i.e. other
members of a gene family), I
do the following test primarily. That is, I made a southern blot of A. thaliana
genomic DNA digested by
3 different restriction enzymes. Then, I hybridize the original probe (which was
used for screening) and
the probe made from the whole insert (9-17 Kb) of the isolated clones. I
expected that the banding
pattern of the S. blots of the isolated clones to be similar (or include) that
of the S. blot probed with
the screening probe. This did not happened for any of the isolated clones. Any
advice or comment
regarding this problem is greatly appreciated. Thanks. Ali
My E-Mail Address is: malboobi at qucdn.queensu.ca
