E. coli and SDS-PAGE

David Martin x3175 dmartin at crc.ac.uk
Fri May 20 04:00:38 EST 1994


In article <bushnt.1119742558A at usenet.rpi.edu> bushnt at rpi.edu (Timothy Bushnell) writes:
>
>    In an undergraduate lab, we are having the students prepare protein
>profiles of a stationary phase E. coli culture using SDS-PAGE.  Currently,
>we spin 50 mls of culture down and resuspend the pellet in 1 ml of 62.5 mm
>Tris pH 6.8, 2% SDS, 5% B-mercapt. and 10% glycerol.  This is then boiled
>for 5 to 10 minutes and the samples then applied to a SDS-PAGE gel.
>
>    The problem is that the sample is very viscous and near impossible to
>successfully load on a gel.  Typically, after the sample is loaded into the
>wells, and the pipet tip removed, the sample sticks to the tip and is
>removed as a gob into the upper tank buffer.  

This sticky stuff is called DNA which, as a protein chemist, I avoid at all
costs.. :-)

Try sonicating the tube for 2 x 10 secs. This normally does the trick for me.

hope this helps.

....David Martin
dmartin at hgmp.mrc.ac.uk




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