Bacillus mucilaginosus Krassilnikov|BNCC335819 |BNCC

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Bacillus mucilaginosus Krassilnikov

Culture medium	Nitrogen fixing medium (ABM):KH2PO4 0.2g,K2HPO4 0.8g,MgSO4 · 7H2O 0.2g,CaSO4 · 2H2O 0.1g,FeCl3 trace g,Na2MoO4 · 2H2O trace g, yeast extract 0.5g, mannitol 20.0g, agar 15.0g, distilled water 1.0L,pH 7.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps; (3) draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly; (4) inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates; ⑤ Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Growth conditions	30 ℃;24-48h; Aerobic;
Storage conditions	2-8 ℃
morphology	Colony diameter is 6-8mm, round, irregular edge, transparent, front white, light color, middle convex, smooth and bright surface, moist texture, easy to pick up, no pigment, G + (blue-purple), bacilli, purity: pure
application	Bacterial fertilizer
Sharing mode	Public welfare sharing

Bacillus mucilaginosus Krassilnikov

Storage conditions : 2~8 ℃

No. 335819

Product format: freeze dried, 200ul

Validity : 6 years

Biosafety level : 1 , handle in ultra-clean table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions :30 ℃, aerobic, azotobacter medium, 24-48h. Azotobacter culture medium: KH2PO4 0.2g,K2HPO4 0.8g,MgSO4 ·7H2O 0.2g, CaSO4·2H2O 0.1g,FeCl3 trace g,Na2MoO4·2H2O trace g, yeast extract 0.5g, mannitol 20.0g, agar 15.0g (not included in liquid medium), distilled water 1.0L,pH 7.2. Sterilization at 121 ℃ for 15min.

Recovery steps:
(1) Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;
(2) Open the ampoule in the biosafety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps;
(3) Draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly;
(4) Inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates;
(5) Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability, in 24-48h liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	Size: 2-4mm Shape: Round Edge: Neat Transparency: Transparent Color: off-white uplift: middle raised surface: bright and smooth texture: moist and viscous
Conclusion:	good viability, no abnormal colony morphology, qualified

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