Clostridium tyrobutyricum|BNCC336696 |BNCC

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Clostridium tyrobutyricum

Culture medium	Enhanced Clostridium agar medium (RCM): yeast extract 3.0g, beef extract 10.0g, glucose 5.0g, tryptone 10.0g, sodium chloride 5.0g, soluble starch 1.0g, cystine hydrochloride 0.5g, sodium acetate 3.0g, agar 20g,pH 6.8±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety 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.
Growth conditions	37 ℃;48-72h.; Anaerobic
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, opaque, gray-white on the front, gray-white on the back, convex in the middle, smooth and bright surface, moist texture, easy to stir up, G + (blue-purple), bacilli, purity: pure
application	butyric acid production
Sharing mode	Public welfare sharing

Clostridium tyrobutyricum

No. : 336696

Storage conditions : 2~8 ℃

Product Format: freeze dried, 200ul

Validity period: 6 years

Biosafety level : 1,handle in ultra-clean table or 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 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 lose of viability.

Growth conditions :37 ℃, anaerobic, Clostridium sporogenes medium, 48-72h. Clostridium sporogenes culture medium: tryptone 10.0g beef paste 10.0g glucose 5.0g sodium chloride 5.0g yeast paste 3.0g sodium acetate 3.0g soluble starch 1.0g cysteine 0.5g agar 0.5g pH6.8±0.2, distilled water 1000ml, 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 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 24-48 hours.

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

item	test results
viability	good viability, in 48-72h liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology: (above)	size: 1-2mm shape: round edge: neat transparency: opaque color: light yellow uplift: middle raised surface: bright and smooth texture: moist and viscous
conclusion	good viability, no abnormal colony morphology, qualified

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