I'm trying to prepare R2B/R2A media with seawater to cultivate marine bacteria after autoclaving (110°C) my media. I systematically observe a important precipitate. Despite many tests (in particular at different percentage of seawater and different types of filtration, I did not succeded to find the solution. Do you have any idea to help me ?
Thank for you help !
One of the major reason of ppt is aggregation of calcium and phosphate. The traditional way is that, prepare with 250ml of DW and autoclave and mix with filter sterilized seawater before pouring.
Kae Kyoung Kwon
Thank you so much for your answer. I tried a similar method (R2B 10X autoclaved then mixed with autoclaved SW), it did not precipitate!
The problem is that to start my cultures, I would like to autoclave the medium directly in glass tubes to avoid maximum contamination. I still haven't found a solution to autoclave the medium in glass tubes.
I also tried letting the bottle precipitate, then autoclaving it and filtering it on 0.2µm and autoclaving it again in the glass tubes. The precipitate problem was solved but it certainly changes the composition of the medium too much.
If you have any ideas I'm interested! Thank you for your help!
Ah~ You trying not easy process! If you will not cultivate all marine microorganisms, prepare media only with NaCl is a way. Actually many marine microorganisms require salts such as Mg, K, or Ca as well as Na. Hence, another way is prepare medium with Na, K, Mg and trace elements solution (If possible to neglect strains require Calcium).
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