Clostridium difficile|BNCC340237 |BNCC

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Clostridium difficile

Culture medium	Liquid thioglycolate medium (FT): 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, 1.0L of distilled water and 7.1±0.2 of pH value. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a φ18mm test tube of 10ml liquid medium ( put in an anaerobic environment for 24h before handling); (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) Place the liquid test tube under the above-mentioned specified growth conditions, and the bacterial liquid is turbid or there is a large amount of bacterial growth at the bottom.
Growth conditions	37 ℃;24-48 hours; Anaerobic
Storage conditions	2-8 ℃
morphology	Clostridium difficile, for Gram-positive bacilli, thioglycolate liquid culture medium culture bacteria liquid turbid, precipitation, it is recommended to use liquid culture.
application	Research, quality control, "GB 4789.28 medium and reagent quality requirements" standard strain
Sharing mode	Public welfare sharing

Clostridium difficile

Storage conditions: 2~8 ℃

No. 340237

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level: 2, handle in safety cabinet

Notes: 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°C, 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, resate Green 0.001g, agar 0.75g, distilled water 1.0L, pH 7.1 ± 0.2. Sterilize at 121°C for 15min.

Recovery steps:

(1)Prepare a φ18mm test tube of 10ml liquid medium; and two agar plates（place in anaerobic environment for 24hours before usage）；

(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 results
viability	good viability, in 24-48h liquid medium become turbid
colony morphology: (above)	Clostridium difficile is a gram-negative bacterium, an oval bacterium that does not produce spores. The liquid culture medium of thioglycolate is turbid. It is recommended to use liquid culture.
Conclusion	good viability, no abnormal colony morphology, qualified