Help! Panning

tokio nakane tnakane at GIPAC.SHINSHU-U.AC.JP
Mon Aug 7 21:29:28 EST 1995

We are currently trying to clone a G protein-coupled receptor by panning method.
We constructed a rat brain cDNA library (pME18S vector; SR-alpha promoter)
and transfected the library to COS-7 cells by DEAE dextran method.  After
72 hrs, we could not detach cells from dishes by the treatment with PBS and
5 mM EDTA at 37 C for 30 min.  The untransfected cells were easily detached
by the treatment.  Any advice would be much appreciated.


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