Lactobacillus plantarum subspecies|361467 |BNCC

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Lactobacillus plantarum subsp. plantarum

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 ℃; 18-24h; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 0.5-1mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, bright and smooth on the surface, moist texture, easy to pick up, G+(blue purple), rod-shaped
Sharing mode	Public welfare sharing

Lactobacillus plantarum subsp.

Storage conditions: 2~8 ℃

No. 361467

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℃, aerobic, 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 a test tube containing 5-10mL of liquid culture medium and 2 plates;

(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 24 hours, plate bacteria layer is obvious; colony is typical on the streaked plate
Colony morphology: (pictured above)	Size: 0.5-1mm Shape: Round Edge: Neat Transparency: Opaque Color: White Protuberance: Middle convex surface: smooth texture: moist and easy to stir
conclusion: good viability, no abnormal colony morphology, qualified

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