Growing BL21 DE3 (pET 9d) Transformants in M9 Minimal Media.

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

[miha250977]

Originally Posted by youg raj

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

I have the same problem...can somebody help, pls?