Purifying a trancripxn factor using biotinylated DNA

David Wong cookie at casbah.acns.nwu.edu
Wed Nov 2 18:16:37 EST 1994


Recently we've found a cell line to provide a source for isolating and
purifying a transcription factor (identified through gel shift).  

To purify the protein, we're using a biotinylated oligo consisting of the
binding region.  We hope then to complex the factor-DNA with streptavadin and
precipitate it with biotinylated resin.  We're using the protocol right out of
Current Protocols.

However, we're having difficulty getting even the DNA to come down with the
resin (the DNA is radiolabelled for tracking purposes).

I was wondering if anyone has had any previous experience using this or a
similar protocol and could offer tips or advice as to what might be going
wrong.  Any additional suggestions in using this method as a whole would also
be appreciated.

Please e-mail me directly.

Thanks.

David Wong
e-mail: davidwong at nwu.edu





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