Lactobacillus fermentum|BNCC194390 |BNCC

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Lactobacillus fermentum

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; Micro aerobic (3%-5% O2,10% CO2)
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, milky white on the front, milky white on the back, convex in the middle, smooth and bright surface, moist texture, easy to stir up, G + (blue-purple), bacilli, purity: pure
Separation substrate	Yogurt
Sharing mode	Public welfare sharing

Lactobacillus fermentum

BNCC No.： 194390

Storage conditions: 2~8 ℃

Product format： freeze dried, 200ul

Validity period: Freeze-dried tube for 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 ℃, slightly aerobic, MRS medium: cheese peptone 10.0g, beef powder 8.0g, 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 0.05g, Tween 80 1.0g, distilled water 1000mL. pH6.2±0.2. 121 ℃,15min.

Recovery steps:

(1) Prepare 2 MRS agar plates

(2) Open the ampoule tube 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) Draw 0.5mL of liquid culture medium into the freeze dried ampoule,make the strain pellet fully dissolved and inoculate a plate with the solution, then distribute it well in 200ml/plate;

(4) Put the agar plates under the above culture conditions for cultivation 24-48 hours.

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

item	test results
viability:	good viability, in 40h MRS plate colony obviously
colony morphology:	size: general color: milky white shape: round edge: neat edge wet and dry: wet and smooth: smooth and bright transparency: opaque bulge: bulge
conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified