Northern question

Cres Bookstein cbookste at nospammedicine.bsd.uchicago.edu
Thu Dec 11 17:26:54 EST 1997


On 10 Dec 1997, Susan Hubchak wrote:  Does anyone know of a way to increase
the stringency of the washes where you won't see cross hybridization, or is
this a pipe dream?
   Our experience with two very closely conserved isoforms was that only
isoform specific probes successfully differentiated the expression of the
two.  We tried extremely stringent hyb and wash conditions (both temp and
salt and single strand digestion after all).  The bottom line was that
whenever full length sequences of either probe were used, the patterns were
identical.  We resorted to short unique fragments on Northerns and RPAs to
determine their true expression patterns.

Perhaps this will save your coworker a lot of work.

Cres Bookstein





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