Hello,
I have grown anaerobic bacteria for years in a coy chamber. I would make anoxic media for years by passing media into the chanber, loosen the cap and let it deoxigenate for 3 days. how does this compare to the hungate or the Wolfe method of making anoxic media? Does the method I have used insufficient to deoxygenate the media?
For me, I use specific kit that provides the required atmosphere for the bacteria that I want to grow them and the plates inserted inside the anaerobic jar with the specific atmospheric kit.
I don’t think it is such a good idea to leave medium with the cap open for three days, as it may get contaminated. Preferably, the solid ingredients should be introduced into the anaerobic chamber and dissolved with deoxygenated water, prior to adding a solution of oxygen scavenger, e.g. cysteine, also made with deoxygenated water, and sterilizing. Anaerobiosis can be visualized with resazurin, which is colorless when properly reduced and pink or violet when oxidize.
Hi,
The method you have described above is fine overall, and that must have been confirmed by the fact that you have got desired growth of your strict anaerobic microbe...I have also used a Coy chamber, so I would like to share an additional point...after transferring the media inside the chamber I left it there for 24 h and after that I have taken the media in a wide mouth beaker and stirred it for 2h atleast using magnetic stirrer set up inside the glove box at about 400-500 r.p.m. to mix it with the anaerobic environment...after this I have used the media for culturing and have got good results... You can try this step and see what differece it is making...the same thing can be done in the last step of preparing deoxygenated water for dissolving dry media components as mentioned by Mr. Beguin above.
Regards,
Shiladitya
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