Rhizobium genus|196223 |BNCC

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Rhizobium sp.

Subculture procedure	① Prepare 1-2 of the above plates; ② After disinfecting the surface of the ampoule tube, open it in a safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ Cultivate the agar plate under the above cultivation conditions for 18-24 hours. The colony diameter is 1-2mm, round, with neat edges, opaque, pink on the front, raised in the middle, smooth and bright on the surface, moist texture, easy to pick up, G - (red), Bacillus, purity: pure, producing mucus
Growth conditions	30 ℃, 18-24h, aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colonies are 1-2 mm in diameter, circular with an entire margin, and opaque. The colony surface is pink, convex, smooth, shiny, and moist in texture. They are easily picked up. The cells are Gram-negative (red), rod-shaped (bacilli). The culture is pure and exhibits mucoid production.
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Rhizobium sp.

Storage conditions : 2~8 ℃

No. : 196223

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 plate 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, rhizobia YMA medium, 18-24h. Rhizobium YMA medium: KH2PO4 0.25g,K2HPO4 0.25g,MgSO4·7 H2O 0.2g,NaCl 0.1g, yeast extract 0.8g, mannitol 10.0g, agar 18.0g, distilled water 1.0 L,pH 7.2,121 ℃,15min sterilization.

Recovery steps:
(1) Prepare 1-2 of above mentioned plates;
(2) Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip in a flame, quickly drop sterile water to creak it, then break it with forceps;
(3) Draw 0.5ml of sterile water into the freeze dried ampoule, make it fully dissolved, and distribute the solution to the plates well in 200ul/plate;
(4) Put the plates under the above culture conditions for cultivation for 18-24 hours.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability, in 24 hours liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	Size: 1-2mm Shape: Round Edge: Neat Transparency: Translucent Color: Pink Protuberance: Middle Protuberance Surface: Bright and Smooth Texture: Wet and Sticky
Conclusion:	good viability, no abnormal colony morphology, qualified

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