Clostridium perfringens|317723 |BNCC

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Clostridium perfringens (Veillon and Zuber) Hauduroy et al.

Subculture procedure	①Prepare 1-2 fresh agar plates (placed in an anaerobic environment for 24 hours of deoxygenation) with typical bacterial colonies; 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 liquid culture medium or sterile water (placed in an anaerobic environment for 24 hours of deoxygenation in advance) and transfer it into a freeze-drying tube, thoroughly dissolve and mix; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Cultivate under the above conditions, and the strain can be used once it grows.
Growth conditions	37 ℃; 24-48 hours; anaerobic;
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony has a diameter of 1-2mm, irregular shape, irregular edges, opaque, gray white front, raised middle, smooth surface, moist texture, easy to pick up, G+(blue purple), Bacillus subtilis
Sharing mode	Public welfare sharing

Clostridium perfringens

Storage conditions: 2~8 ℃

No.: 317723

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. 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, thioglycolate liquid medium, 24-48h. Thioglycolate liquid medium: caseptone (trypsin hydrolysis) 15.0g, yeast extract 5.0g, glucose 5.0g, sodium thioglycolate 0.5g,L-cystine 0.5g, sodium chloride 2.5g, azure 0.001g, agar 0.75g, distilled water 1.0L,pH 7.1 ± 0.2. Sterilization at 121 ℃ for 15min.

Recovery steps:
(1)Prepare a φ18mm test tube of 10ml liquid medium;

(2)Sterilizing the ampoule, open it in 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 the freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly;

(4)Put the test tube under the above culture conditions for cultivation. The culture solution is obviously turbid or mass growth at the solution bottom occur, this indicates the bacterial grows well.

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

item	test result
viability	good viability,in 24-48h liquid medium become turbid
colony morphology: (above)	Clostridium perfringens is a gram-positive bacterium. The bacterial liquid of thioglycolate liquid culture medium is turbid, and there is a large amount of bacterial precipitation at the bottom. Liquid culture is recommended.
Conclusion	good viability, no abnormal colony morphology, qualified

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