growth of E.coli in minimal media(m9) is very less
Posted 16 January 2011 - 12:47 AM
I m trying to grow my wild type e.coli strain(ATCC 27235) in m9 media with 2mg/ml of glucose. I had made the media as given
1.Dissolve Disodium Phosphate (anhydrous) 33.9 g ,Monopotassium Phosphate 15.0 g, Sodium Chloride 2.5 g ,Ammonium Chloride. 5.0 g In 1 L of purified water.
2. Autoclave at 121°C for 15 minutes.
3. To prepare M9 Minimal Salts Medium, add 200 mL sterileM9 Minimal Salts, 5Ũ to 750 mL sterile purified watercooled to 45-50°C, adjusting the final volume to 1 liter.
4. Aseptically add 20 mL filter-sterilized 20% glucose solution, 2 mL sterile 1.0 M MgSO4 solution and 0.1 mL sterile 1.0 M CaCl2 solution. Mix well. Adjust ph 7.4 with NaOH
But the growth of E.coli is very less (O.D. after 16 hours=0.147).after that o.d. decreases
I read few papers I which people have talked about minimal media should be supplemented(with thiamine or feso4 etc etc)
So my question is that For geting more growth of Wild type E.coli on m9 media with 2mg/ml of glucose on recipe as given above is it compulsory to add supplements(which?) or I can get growth without that?
Posted 16 January 2011 - 11:18 PM
Posted 17 January 2011 - 04:27 AM
I got this strain from NCIM lab and it is their mistake.
I will try that one.
even I got few papers on ATCC 27325growing on m9
Thanks for your suggestions