chloroform in colony blots

Jeff E. Janes jejanes at silibone.cchem.berkeley.edu
Wed Nov 25 14:21:25 EST 1998


Hi all,

  I'm doing some experiments in which I lift colonies onto 
nitrocellulose, then visual for a specific protein using 
antibody-AP conjugate.  Some of the protocols I have seen
(none of which seem quite satisfactory for me) call for treating
the nitrocellulose-bound cells (E. coli) to chloroform vapors
for 15 minutes prior to lysozyme treatment.  Does anyone know
the purpose of this step?

Also, I would prefer to blot for periplamsic proteins only,
but all protocols indicate complete cell lysis.  Does anyone
know of a protocol which will work for this?

Thanks,

Jeff



More information about the Methods mailing list