Anabaena variabilis
ATCC® 29413 BioVector NTCC质粒载体菌种细胞基因保藏中心

Anabaena variabilis
ATCC® 29413 BioVector NTCC质粒载体菌种细胞基因保藏中心 Designation: [IUCC 1444; MSU A37]
Deposited Name: Anabaena variabilis Kutzing
Medium
ATCC® Medium 616: Medium BG11
for bluegreen
algae
Growth Conditions
Temperature: 26.0°C
Atmosphere: Under light intensity of 2,0003,000
lux
Propagation Procedure
1. Open vial according to enclosed instructions.
2. Using a single tube of #616 broth (5 to 6 ml), withdraw approximately 0.5 to 1.0 ml with a Pasteur or 1.0 ml
pipette. Rehydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a second tube of broth and a #616 slant.
5. Incubate tubes at 26oC under 20003000
LUX light. It is helpful to incubate test tubes in a
slanted position to increase gas exchange in broth and to enhance exposure to light.
Good growth, indicated by increased pigmentation in the broth or on the slant, should occur after two weeks
of incubation. Examine cells microscopically to assure that they are intact and healthy. At this time additional
test tubes or flasks can be inoculated. A 5% inoculum is recommended (i.e. 5 ml of culture to 100 ml fresh
medium).
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.
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