E. coli and SDS-PAGE

Eric R. Hugo e_hugo at dsu1.dsu.nodak.edu
Sat May 21 07:38:47 EST 1994


I agree with Virginia in that it sound like you have way too much cellular 
material in your sample.  Typically when I used to look at E. coli raw samples 
on minigels (10 x 8 cm) I usually tried to load approx. 0.04 OD 550 nm units 
of cell culture per lane.  Generally this worked out to putting 0.2 OD units 
worth of culture (eg 10 ul of an OD 20 stationary phase broth culture + 10 ul 
5X loading buffer + 30 ul water) in a microfuge tube, boiling, 30 sec spin in 
a microfuge, and then loading 10 ul / lane.  This typically gave me plenty of 
signal on a Coomassie-stained gel.  If you go to silver staining the amounts 
can be reduced significantly.
				Eric 



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