[Protein-analysis] Growing BL21 DE3 (pET 9d) Transformants in M9 Minimal Media.

youg raj via proteins%40net.bio.net (by yougraj from gmail.com)
Mon Oct 23 06:34:42 EST 2006


Hi to everyone,
I have question regarding the protocol for growing cells in M9 media, to
express N15 labelled protein.
Cells are growing nicely in LB but when I am inocluting them @1ml/500ml in
Minimal media of  50ml overnight culture resuspended in 10ml M9, cells are
not growing at all. I am adding all trace elements, and traditional receipe
for salts. Glucose in 10g/L.
Initially OD is approx. 0.6 at 600nm. Cells are always grown @37 degrees.
Any suggestions plz.
(other people in my lab using the same protocol and strain are growing and
expressing protein nicely).
-- 
Youg Raj
A/P G. Grubers Lab's
School of Biological Sciences
NTU, Singapore 637551
hp:  +65-8133 8050
Lab.+65-63162852


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