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Minimal media

Tom Dougherty Doughert at synapse.bms.com
Mon Nov 4 11:45:12 EST 1996


In article <327D779F.2991 at sluvca.slu.edu>, "Joseph E. Mayer Jr." <mayerje at sluvca.slu.edu> says:
>
>Haishan Xiong wrote:
>> 
>> I have been experiencing some problems with growing E. coli in M9
>> minimum media.  I tried to grow JM109 and XL1-Blue cells in M9 +
>> glucose.  Neither grew.  I did not find any difficiency in terms of
>> their genotype.  How can I get around with this problem?
>> I appreciate your kindly help.
>
Have you added thiamine HCl?  It is necessary to have this vitamin present
in low concentration (0.5 ug/ml, final). It also helps to culture your E.
coli cells on M9 agar plates before going into M9 liquid culture.  Solidify 
M9 glucose with 1.75% Noble agar.  They usually take 48 hrs to adapt to the 
minimal media after coming off LB or other rich nutrient sources. Finally, 
add sodium bicarbonate to liquid cultures in M9 glucose at 400 ug/ml final
concentration. The bicarbonate must be filter sterilized, not autoclaved.
 
Hope this helps,
   Tom Dougherty



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