Anabaena doliolum Bharadwaja
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
1. Withdraw 0.6 ml from the base of a broth culture where cells are concentrated, or harvest cells from a slant culture with 0.6 ml of #616 broth.
2. Using this aliquot, inoculate one broth and one slant tube with 0.2 and 0.4 ml respectively.
3. Incubate tubes at 26oC under 2000-3000 LUX light.
To minimize change in a culture, it is recommended that a frozen seed stock be established from early passage cells. This may be accomplished by propagating the strain under ideal conditions, utilizing recommended medium, temperature and light. Prepare a concentrated cell suspension, after good growth is achieved. If grown in broth, pellet the cells by centrifugation. Decant the supernatant and resuspend the pellet in fresh #616 broth using 1/10 or less of the original volume. For slant cultures, wash cells off the agar surface with a minimal amount of #616 broth so that a concentrated cell suspension is attained. Add 50% DMSO solution to the concentrated cell suspension so that the final concentration of DMSO in the suspension is 5%. Dispense small aliquots (0.5 to 1 ml) of the suspension into small sterile vials. Store the vials at -50oC or below.
When needed, remove vials from storage, thaw contents in a 37oC water bath and inoculate into recommended medium. A minimum of 0.2 ml of the thawed stock should be used to inoculate 5 ml of broth or 1 agar slant.
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