Clostridium perfringens toxin type A|125404 |BNCC

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Clostridium perfringens Type A

Subculture procedure	① Prepare 1-2 of the above 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 transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into two plates, each Agar plate containing about 200 μ L, and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	37 ℃, 18-24h, anaerobic
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony has a diameter of 1-2 mm, with an irregular shape and uneven edges. It is opaque, flat in morphology, and appears grayish-white on the front side, with a dull and moist surface. It is Gram-positive (staining blue-violet) and consists of rod-shaped bacteria.
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Clostridium perfringens Type A

Storage conditions: 2~8 ℃

No.: 125404

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level : 2, handle in safety cabinet operation

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 medium: casepone (trypsin hydrolysis) 15.0g, yeast extract 5.0g, glucose 5.0g, sodium thioglycolate 0.5g,L-cystine 0.5g, sodium chloride 2.5g, azurite 0.001g, agar 0.75g, 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 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 18-24h liquid medium beccome turbid, plate colonies are obvious
colony morphology: (above)	Size: 2-4mm Shape: Round Edge: Irregular Transparency: Opaque Color: white uplift: middle raised surface: bright and smooth texture: moist and viscous
Conclusion:	good viability, no abnormal colony morphology, qualified

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