2 replies to this topic

[ddk]

hi every one,

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, 5× 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?
Thanks

[protolder]

Hola, I check ATCC number 27235 and it isn´t an E. coli strain. Anyway you have to check the genotype of your stain and´you have to add all the compound that it is´t able to sintetize an thiamine or proline, or whatever. if you want grow more and it hasn´t any aminoacid auxotrophy you can add casaminoacids and tryptophan, because the basal M9 formula has low nitogen proportion. Buena suerte

[ddk]

I am sorry that number is 27325,
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 alot   


Thanks for your suggestions

protolder, on 16 January 2011 - 11:18 PM, said:

Hola, I check ATCC number 27235 and it isn´t an E. coli strain. Anyway you have to check the genotype of your stain and´you have to add all the compound that it is´t able to sintetize an thiamine or proline, or whatever. if you want grow more and it hasn´t any aminoacid auxotrophy you can add casaminoacids and tryptophan, because the basal M9 formula has low nitogen proportion. Buena suerte