Sake Lactobacillus subsp. sake|192620 |BNCC

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Lactobacillus sakei subsp. sakei

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

Lactobacillus sakei subsp. sakei

Storage conditions : 2~8 ℃

No. : 192620

Product format: freeze dried, 200ul

Validity : 6 years

Biosafety level: 1 , handle in ultra-clean table or safety cabinet

Notes: 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 ℃, aerobic, 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. 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 results
viability:	good viability, in 24-48h the liquid medium become turbid, and the plate strain is weak.
colony morphology: (above)	Lactobacillus sake subspecies, Gram-positive bacilli, MRS liquid culture bacterial fluid turbid, There is a large amount of precipitation at the bottom of the test tube, and the growth of the plate strain is weak. Liquid culture is recommended.
Conclusion:	good viability, no abnormal colony morphology, qualified

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