Lactobacillus johnsonii|BNCC186384 |BNCC

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Lactobacillus johnsonii

Culture medium	MRS medium (MRS): 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.
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 ℃;18-24h; Anaerobic;
Storage conditions	2-8 ℃
morphology	Size: 0.5-1mm shape: round edge: neat transparency: opaque Color: milky white bulge: middle bulge surface: bright and smooth texture: moist and easy to pick
Separation substrate	Human blood
Sharing mode	Public welfare sharing

Lactobacillus johnsonii

Storage conditions: 2~8 ℃

No. 186384

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 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℃, anaerobic, MRS medium, 18-24h, MRS medium: peptone 10.0g, beef extract 10.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0 g, 2.0 g of dipotassium hydrogen phosphate, 0.04 g of manganese sulfate, 1.0 g of Tween 80, 20.0 g of agar (not included in liquid medium), and 1.0 L of distilled water. pH 5.7±0.2. Sterilize at 121℃ for 15min.

Recovery steps:

① Prepare 1 test tube containing 5~10mL liquid medium and 2 plates (placed in an anaerobic environment to deoxygenate for 24h in advance)

② Open the ampoule 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.

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

Item	test results
viability:	good viability, in 18-24hours liquid medium become turbid,strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	size: 0.5-1mm ； Shape: round ; Edge: neat ; Transparency: opaque color: milky white; Protuberance: middle bulge ; Surface: bright and smooth ; Texture: moist and easy to stir
Conclusion	good viability, no abnormal colony morphology, qualified