Clostridium ramosum|BNCC359261 |BNCC

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Clostridium ramosum

Culture medium	Columbia blood plate (English name: CA-B): casein pancreatin digest 10.0g, cardiopancreatin digest 3.0g, corn starch 1.0g, meat gastric enzyme digest 5.0g, yeast extract 5.0g, sodium chloride 5.0, agar 20.0g, distilled water 1.0L,pH 7.3±0.2. After sterilization at 121 ℃ for 15min, cool to 55 ℃, add 5% sterile defibrated sheep blood, shake well, pour the plate and cool it for later use.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it 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.
Growth conditions	37°C;24-48h; Anaerobic
Storage conditions	2-8 ℃
morphology	Size: 0.5-1mm shape: round edge: neat transparency: opaque color: off-white bulge: middle bulge surface: bright and smooth texture: moist and easy to stir up, G + (blue-purple), bacilli, purity: pure
Sharing mode	Public welfare sharing

Clostridium multibranch

Storage conditions : 2~8 ℃

No. : 359261

Product format :freeze dried, 200ul

Validity : 6 years

Biosafety level : 2, handle in 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: 37 ℃,5% CO₂, Columbia blood plate (ready-to-use type),18-24h.

Recovery steps:

① Prepare the above flat plate 1-2 block (put it in anaerobic environment for 24 hours in advance);

② Open it in the safety cabinet, burn the top with an alcohol lamp, quickly drop sterile water to break it, and then break it with tweezers;

③ suck 0.5mL of sterile water and put it into the freeze-drying tube. after fully dissolving the bacterial powder, put it into the plate 200 μL/piece, coated evenly;

④ Place the plate under the above culture conditions, and the strain can be used when it grows out.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability	good viability，in 48hours strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	size: 0.5-1mm shape: round edge: neat transparency: opaque Color: off-white uplift: middle convex surface: bright and smooth texture: moist and easy to stir
Conclusion	good viability, no abnormal colony morphology, qualified

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