I recently received a live culture of Sulfolobus shibatae which came with a separate vessel of pre-prepared media. I also made media following the attached recipe. I immediately began culturing the cells at 76C, 230 RPM in both the pre-prepared media and my media.
After 4-5 days of culture, cell density increased in both the pre-prepared media, and my media.
I then re-inoculatied cells into fresh media (both types), however, It has been 5 days since inoculation and I cannot see growth in either my media, or the pre-prepared.
Has anyone experienced such a problem before when growing Sulfolobus? Is the media stable, or does it spoil very quickly? I understand Na2MoO4 is light sensitive, however, the my media was freshly prepared upon inoculation, so I cannot see this being the cause for no growth.
Thanks in advance!
Keep the T-25 flask in dark, stabilize the media at least for 96 hrs you will find growth. Or your media is lacking some cell growth and colony stimulating factors.
To follow up on this. Culturing in falcon tubes appears to have been the cause of the problem. The falcon tube degraded slowly, resulting in contaminant in the media - seen under EM. This appears to have inhibited growth, as culture in T-25 (or polystyrene container) removed contaminant and allowed higher cell density.
Thanks for help!
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