Liquid B-gal assays???? How to?

Marieke R. Koedood Zhao rkoedood at bio.bu.edu
Mon Dec 16 13:16:16 EST 1996


We also use Z-buffer and ONPG, but lyze the cells by 5 fast freeze-thaw
cycles.

Marieke Koedood Zhao


Rachael Buchnik (rbuchnik at MAIL.NCIFCRF.GOV) wrote:

: you can use the protocol published in FEMS microbiol lett 128 (1995) 
: 201-206 by Fred Kippert.
: 1. To 10-200 micro litters of cells you add 600 microliters of Z buffer 
: with 0.2% Sarcosyl (Sodium Lauroyl Sarcosinate). 
: 2. Incubate for at least 30 minutes.  
: 3. Add 150 microliters of ONPG 4 mg/ml.
: 4. You can stop the reaction by adding 400 microliters of Na2CO3 1.5M.

: Thats all.  Simple and quick.
: Good luck!

: On Thu, 12 Dec 1996 mike at ncsu.edu wrote:

: > I would appreciate your sharing your favorite protocol for liquid
: > B-gal assays with me.  I am using PJ69.4a for a 2-hybrid screen and
: > will potentially being doing many at one time (or hopefully a few).  I
: > have tried grinding the cells with glass beads and then incubating the
: > extract with X-gal and Z buffer but it didn't work out (have yet to
: > repeat this).  Just wondering if anyone knows of a good way to do
: > this.
: > 
: > Thanks,
: > Mike Faughn
: > 
: > Dept. Genetics, NCSU
: > 
: > 
: > 



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