Clostridium perfringens|251435 |BNCC

Essential Information Certificate Related Products

Enterocloster bolteae

Subculture procedure	① Dissolve the frozen tube in a 37 ℃ water bath and shake quickly to dissolve; ② Wipe the outer wall of the Frozen vial with an alcohol swab for disinfection, and then transfer it to a safe cabinet for operation; ③ Unscrew the tube cap, aspirate all the dissolved solution, and transfer it into 1-2 agar plates (each Agar plate contains approximately 200 μ L, and for anaerobic bacteria cultivation, the culture medium needs to be placed in an anaerobic environment for 24 hours to remove oxygen); ④ After uniform coating, transfer to the above cultivation conditions for cultivation.
Growth conditions	37 ℃; 48-72 hours; anaerobic
Storage conditions	-80 ℃
Safety level	2
morphology	The colony diameter is 0.5-1mm, circular, with neat edges, gray white on the front, raised in the middle, smooth surface, bright surface, moist texture, easy to pick up, G+(blue purple), Clostridium, purity: pure
Sharing mode	Public welfare sharing

Clostridium bolteae

Storage conditions: 2~8 ℃

No. 251435

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 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, improved GAM medium, 2-3d, improved GAM medium: peptone 5.0g, chicum 5.0g, soybean peptone 3.0g, yeast powder 2.5g, beef powder 2.2g, digested serum powder 10.0g, beef liver powder 1.2g, glucose 0.5g, potassium dihydrogen phosphate 2.5g, sodium chloride 3.0g, soluble starch 5.0g,L-cysteine 0.3g,L-arginine 1.0g,L-tryptophan 0.2g, sodium thioglycolate 0.3g,PH7.3±0.1,121 ℃,15min.

Recovery steps:
(1) Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates(put in an anaerobic environment for deoxygenation in advance for 24 hours);
(2) Open the ampoule 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.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability, in 48h strain layer is obvious; colony is typical on the scribed plate
colony morphology: (above)	size: 2-4mm shape: round edge: irregular transparency: opaque color: light yellow uplift: middle convex surface: bright and smooth texture: moist
Conclusion:	good viability, no abnormal colony morphology, qualified

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