Clostridium sporogenes|186188 |BNCC

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Clostridium sporogenes

Subculture procedure	① Prepare 2 plates; ② Open the 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 pour it into the freeze-drying tube. After fully dissolving, mix well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all the plates under the above cultivation conditions, and once the strains grow, they can be used.
Growth conditions	37 ℃; 2-3 days; anaerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	Colony diameter: 1–2 mm, circular, irregular margin, opaque, grayish-white on the obverse, convex in the center, smooth and shiny surface, moist texture, easily picked, Gram‑positive (bluish‑purple), bacillus, purity: pure.
Sharing mode	Public welfare sharing

Clostridium beijerinckii

No. 186188

Storage conditions: 2~8 ℃

Product format: freeze dried,200ul

Validity period: 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:37 ℃, anaerobic, 2-3 days, ① Columbia blood plate (ready-to-use type) ② liquid thioglycolate culture medium: 15.0g of caseptone (trypsin hydrolysis), 5.0g of yeast extract, 5.0g of glucose, 0.5g of sodium thioglycolate, 0.5g of L-cystine, 2.5g of sodium chloride, 0.001g of azure, 0.75g of agar, distilled water 1.0L,pH 7.1±0.2. Sterilization at 121 ℃ for 15min.

Recovery steps:

(1)Prepare sterile test tube of 5-10ml liquid medium and 2 plates; Silica gel plug is used for liquid test tube culture medium to ensure ventilation and facilitate deoxygenation. Please put the liquid medium and plate in an anaerobic environment for 24h before usage.

(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 liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly;

(4) Put the plates under the above culture conditions for cultivation for 48-72 hours.

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

item	test result
viability:	good viability,in 2-3 days strain layer become obvious, colony is typical on the streaked plate
colony morphology: (above)	size: 2-4mm shape: round edge: irregular transparency: opaque color: white bulge: middle bulge surface: rough fold texture: wet and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified

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