symbiotic clostridium|BNCC359144 |BNCC

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symbiotic clostridium

Culture medium	EG medium (EG): beef extract 2.4g, peptone 3 10.0g, yeast extract 5.0g,Na2HPO4 4.0g, glucose 1.5g, soluble starch 0.5g,L-cystine 0.2g, agar 15.0g,L-cysteine HCl H 2 O 0.5g, horse blood 50.0 ml, distilled water 950.0mL, L-cysteine was added to 50 ml 1mol/L HCl and mixed well. Add the remaining components to bring the volume to 950 ml. Adjust the pH to 7.6-7.8. After autoclaving at 121 ℃ for 15min, it was cooled to 50 ℃, and 50.0ml horse blood was added aseptically. Mix thoroughly and pour into a sterile petri dish.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it 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.
Growth conditions	37 ℃;4-5 days; Anaerobic;
Storage conditions	2-8 ℃
morphology	Size: 0.5-1mm shape: round edge: irregular transparency: opaque Color: off-white bulge: middle bulge surface: bright and smooth texture: moist and easy to stir up
Sharing mode	Public welfare sharing

Clostridium symbiotic

Storage conditions : 2~8 ℃

No. : 359144

Product format :freeze dried, 200ul

Validity : 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 of agar plate 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 ℃, strictly anaerobic, EG medium, 4-5 days. EG medium: beef extract 2.4g, peptone 3 10.0g, yeast extract 5.0g,Na₂HPO₄4.0g, glucose 1.5g, soluble starch 0.5g,L-cystine 0.2g, agar 15.0g,L-cysteine HCl·H 2 O 0.5g, horse blood 50.0 mL, 950.0mL of distilled water, add L-cysteine to 50 mL 1mol/L HCl and mix well. Add the remaining components to bring the volume to 950 mL. Adjust the pH to 7.6-7.8. After autoclaving at 121 ℃ for 15min, it was cooled to 50 ℃, and 50.0ml horse blood was added aseptically. Mix thoroughly and pour into a sterile petri dish.

Recovery steps:

① Prepare 2 of the above plates;

② After disinfecting the surface of the ampoule tube, open it in the safety cabinet, burn the top with an alcohol lamp, quickly drop sterile water to break it, and then break it with tweezers;

③ Suck about 0.5mL of sterile water (placed in anaerobic environment for 24 hours) and drive it into a freeze-drying tube. after fully dissolving the bacterial powder, drive it into 2 plates, 200 & mu;L/piece, evenly coated;

④ Place the plate under the above culture conditions for 4-5 days.

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

Item	test results
viability	good viability， in 5days strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	size: 0.5-1mm shape: round edge: irregular transparency: opaque Color: off-white uplift: middle convex surface: bright and smooth texture: moist and easy to stir
Conclusion	good viability, no abnormal colony morphology, qualified

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