Hi,

I could not find any commercially available Hartman's de Bond medium online. We often come accros modified harman's de bond medium in the literature for example: (2004, Shleeva et.al "Formation of ‘non-culturable’ cells of Mycobacterium smegmatis in stationary phase in response to growth under suboptimal conditions and their Rpf-mediated resuscitation "); which show another protocol to prepare the medium.

Another possibility could be to used 10% Dubos medium as a starvation medium. this is what have been done in the multiple stress model of Deb et al. 2009 ("Deb C, Lee C-M, Dubey VS, Daniel J, Abomoelak B, Sirakova TD, et al. A novel in vitro multiple-stress dormancy model for Mycobacterium tuberculosis generates a lipid-loaded, drug-tolerant, dormant pathogen. PLoS ONE nd;4"). you can buy Dubos medium online.

I hope this help, good luck!

best wishes,

Vincent. 

Hello Vincent! Thank you for answer. I just to ask you have you ever used the Hartman's de bond medium? In this case did you prepare all the time fresh mineral stock (because mine passed from grey to pink!)

No, I'm sorry I have never used this medium myself. I do not know why you had this change in colour. Having recently write a review about phenotypes and mycobacteria we came across several papers that used Hartman's de bond medium, in dormancy models for example.

I read this paper from 1999 ("Smeulders MJ, Keer J, Speight RA, Williams HD. Adaptation of Mycobacterium smegmatis to stationary phase. J. Bacteriol. 1999; 181:270–283. [PubMed: 9864340]") where they say: "To prevent precipitation of salts during preparation of this medium, components were added in the listed order..." 

So the order you added you chemical could be an explanation. (They added the full preparation protocol and the order the chemicals where added in their method section). 

hope this help.

Best,

Vincent.  

Actually I followed this paper to prepare the medium, but nothing was mentioned about the change in color! Thank you for you answer!!