How to express toxic proteins in bacteria - correction!
Dr. Peter Gegenheimer
PGegen at UKans.nolospamare.edu
Fri Sep 15 19:39:47 EST 2000
On Sat, 16 Sep 2000 00:21:39, "Emir Khatipov"
<ekhatipo at NOSPAMmidway.uchicago.edu> wrote:
ð The main inconvenience for
ð me is that when the ultracentrifugation is omitted, the lysate appears
ð cloudy, and seemingly contains chelators that cause sheering of cobalt from
ð Talon resin I used to purify His6-proteins. Is there any trick to eliminate
ð ultracentrifugation and the greazy stuff?
I don't recall that we've seen such viscosity with regular (non-His-tagged)
clones. My guess is that the viscosity comes from DNA; if so, the DNA can
normally be sheared to small fragments by squirting the lysate through a 20-
to 22-gauge syringe needle, or brief sonication, or treatment with DNase I.
o----------------------------------------------------------------------o
| Dr. Peter Gegenheimer | Vox: 785-864-3939 FAX: 785-864-5321 |
| Department of | PGegen at UKans.nospam.edu |
| Molecular Biosciences | http://rnaworld.bio.ukans.edu/ |
| University of Kansas |"When you have excluded the impossible, |
| 2045 Haworth Hall | whatever remains, however improbable, |
| Lawrence KS 66045-2106 | must be the truth." S. Holmes |
o_____________________________|________________________________________o
More information about the Proteins
mailing list