Lactobacillus rhamnosus|185356 |BNCC

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Lactobacillus rhamnosus

Subculture procedure	① Prepare one test tube containing 5-10 mL of liquid culture medium and two plates; ② Open the safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Transfer 0.5mL of liquid culture medium into a freeze-drying tube, dissolve it thoroughly, and then transfer it back into the liquid test tube, mixing well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all liquid test tubes and plates under the above cultivation conditions, and the bacterial strains can be used once they grow.
Growth conditions	37 ℃; 24-48 hours; slightly aerobic (3% -5% O2, 10% CO2);
Storage conditions	2-8 ℃
Safety level	1
morphology	Size: 1-2mm Shape: Circular Edge: Neat Transparency: Opaque Color: Milky White Rise Degree: Middle Rise Surface: Bright and Smooth Texture: Moist and easy to pick up
Sharing mode	Public welfare sharing

Lactobacillus rhamnosus

Storage conditions : 2~8 ℃

No. : 185356

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. 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 ℃, slightly aerobic, 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. Sterilization at 121 ℃ for 15min.

Recovery steps:

(1)Prepare a φ18mm test tube of 10ml liquid medium.

(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 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: Irregular Transparency: Opaque Color: milky white uplift: middle raised surface: bright and smooth texture: moist and viscous
Conclusion:	good viability, no abnormal colony morphology, qualified

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