methods for lysis of tissue culture cells

Dr. Jeff Lightfoot jlight at RESUNIX.RI.SICKKIDS.ON.CA
Tue Jun 20 23:38:57 EST 1995


Anybody have a method for lysing small quantities of lymphoblasts or
fibroblasts for SDS-PAGE and Western blotting without boiling?  My main
interest is a procedure that will completely shear DNA.  I've tried
sonication, but our probe isn't that convenient to use because of the small
volumes that I'm using (less than 100 ul).  Use of needle to shear the DNA
works a bit, but there's still a lot of unsheared DNA present.  I'd like to
use DNase, but the lysis buffer that I use has either SDS or urea or both,
which I understand will denature DNase.  Thanks for your help.

Jeff.

"All too often a man's accumulated knowledge dies with him"

        James A. Peters on the life and experiences of L. J. Stadler.






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