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

Rachael Buchnik rbuchnik at MAIL.NCIFCRF.GOV
Sun Dec 15 08:53:29 EST 1996


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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