Clostridium sporogenes|104015 |BNCC

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Clostridium sporogenes

Subculture procedure	① Prepare two of the above-mentioned agar plates and sterile water (pre-equilibrated in an Anaerobic environment for 24 hours);② Disinfect the surface of the ampoule tube, open it in a biosafety cabinet, flame the top with an alcohol lamp, then quickly drip sterile water onto it to crack it open, and use tweezers to break it apart;③ Aspirate 0.5 mL of sterile water, inject it into the freeze-dried tube, fully dissolve the bacterial powder, then transfer it onto two agar plates, 200 μL per plate, and spread evenly;④ Place the plates under the aforementioned culture conditions for incubation. The bacterial strain can be used once growth is observed.
Growth conditions	37 ℃; 18-24h; Anaerobic
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony has a diameter of 0.5–1 mm, appearing circular with neat edges and an opaque texture. It is grayish-white on the obverse side, with a raised center, smooth surface, and moist consistency. The colony is easily picked up, tests Gram-positive (showing blue-violet staining), and consists of rod-shaped bacteria.
Sharing mode	Public welfare sharing

Clostridium sporogenes

Storage conditions : 2~8 ℃

No. : 104015

Product format : freeze dried,200ul

Validity period: 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 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, 18-24h,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. 121 ℃,15min sterilization

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 18-24 hours.

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

item	test result
viability	good viability, in 18-24 hours the liquid medium become turbid, and the plate strain is growing well;
colony morphology: (above)	Clostridium sporogenes, gram-positive bacilli, cultured in liquid thioglycolate medium, turbid bacterial liquid, anaerobic culture of Colombian blood plate growing well.
Conclusion:	good viability, no abnormal colony morphology, qualified

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