Cheese like Lactobacillus|134415 |BNCC

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Lacticaseibacillus paracasei

Subculture procedure	① Prepare 2 of the above plates (placed in an anaerobic environment for 24 hours of equilibration); ② After disinfecting the surface of the ampoule tube, open it in a 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 (placed in an anaerobic environment for 24 hours) and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into 2 plates at a rate of 200 μ L per Agar plate 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 ℃, 24-48 hours, facultative anaerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 0.5-1mm, circular, with neat edges, opaque, white on the front, raised in the middle, smooth on the surface, dry texture, G+(blue purple), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Lactobacillus casei

Storage conditions: 2~8 ℃

No.: 134415

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level : 1, handle in ultra-clear 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 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, MRS medium, 24-48h. MRS medium: peptone 10.0g, beef paste 10.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.04g, Tween 80 1.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L. pH 5.7±0.2. 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 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 24-48 hours.

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

item	test result
viability	good viability, in 24-48h plate bacteria layer is obvious, after marking, colony morphology is typical;
colony morphology: (above)	Size: 1-2mm Shape: Round Edge: Neat Transparency: Opaque Color: milky white bulge: middle bulge surface: bright and smooth texture: moist and viscous
Conclusion:	good viability， no abnormal colony morphology, qualified

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