E.coli dies. HELP!!!!!

Chi-kuang Wen ckwen at expert.cc.purdue.edu
Fri Jun 14 17:23:07 EST 1996


P.R. Alefounder (mbpra at s-crim1.dl.ac.uk) wrote:
: In article <4p6kki$a7t at oravannahka.Helsinki.FI>, kstenber at cc.Helsinki.FI (Kaj Stenberg) writes:
: ...
: |> - all plasmids grown in small volume -> western. No growth problem,
: ...
: |> - All flasks (4) die simultaneously. Pre-ymp 150ml grown, 25ml into each
: |> flask (OD<0.1). OD in production flasks rise to 0.6, then down to zero.
: ...
: |> Obviously medium, temperature, shaking, light, pH, and so on are always
: |> identical.

: Not quite. The flask size, and hence aeration, will be different.

: |> -second idea was phage. I do not think so

Hi,

Are you trying to isolate proteins produced by theE.coli host?
Are the proteins produced toxic to the E.coli?
If the proteins are toxic to the host, you may (1_)culture the E.coli at 
room temp, and (2) and glucose to inhibit the lac promoter activity(I assume
that your plasmid construct is driven by lac). Upon induction, change the
medium which contains no glocose and then add IPTG for a quick induction.

regards,

Chi-Kuang





More information about the Methods mailing list