E. coli growing in M9 minimal media - (Sep/13/2007 )
Hi everyone,
I've began to cultivate E. coli BL21 Codon Plus cells in M9 Media. The media composition is exact as described in Sambrook et al. I also added Thiamine and Biotin to a final concentration of 0,01%. I just would like to check, if the cells also incorporate CoSO4 in my protein, so i added a final concentration of 100 µM to the suspension.
I observe a very very low growth rate; does anyone know how long the lag phase in M9 media is or if CoSo4 could also inhibit or prolong cell growth? I wonder if i figured it out right, but the OD of the cell suspension is showing something different :/
Thanks in advance.
I've began to cultivate E. coli BL21 Codon Plus cells in M9 Media. The media composition is exact as described in Sambrook et al. I also added Thiamine and Biotin to a final concentration of 0,01%. I just would like to check, if the cells also incorporate CoSO4 in my protein, so i added a final concentration of 100 µM to the suspension.
I observe a very very low growth rate; does anyone know how long the lag phase in M9 media is or if CoSo4 could also inhibit or prolong cell growth? I wonder if i figured it out right, but the OD of the cell suspension is showing something different :/
Thanks in advance.
Hi,
I was using M9 Media with BL21 as well. Generally, the cells were growing considerably slower. I don't really know the length of the lag phase, I can just tell you that it takes ages to get them to an OD of even 0.8 (very semi-quantitative statement, I know...
). A standard culture went up to 1.4 in the same time. Also it was hard to rech high OD values at all, my cells just stopped growing at some point...But I also had the impression, that you have to be careful to make sure, the pH of that media is in the correct range. Happens easily that it's is too acidic, which makes things even worse.
I don't know about M9 medium but I do know that 100 µM of Cobalt sulphate in LB is bad news for a culture. At 100µM CoSO4 the E.coli culture grows very very slowly (for all intents and purposes, the culture is a failure). At 10µM of cobalt results in an increased lag time of about 1hr in LB. I can only imagine it is worst for M9. The lag time in M9 is longer then LB. At 1µM Cobalt is stimulatory to E.coli growth. I am now trying value between 1µM and 10µM. But if you do find where the optimum cobalt concentration is let me know. Thanks