Lactobacillus delbrueckii subsp. lactis|289804 |BNCC

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Lactobacillus delbrueckii subsp. lactis

Subculture procedure	① Prepare 1 liquid culture medium with a diameter of 18mm, approximately 10mL; ② Disinfect the surface of the ampoule tube, open it in a safe cabinet, burn the top with an alcohol lamp, quickly add sterile water to the burning area to break it, and then use tweezers to knock it open; ③ Take about 0.5mL of liquid culture medium and transfer it into a freeze-drying tube. After fully dissolving, take it back into the liquid culture medium in the test tube and mix well; ④ Place the liquid test tube under the specified "culture conditions" mentioned above for cultivation. If the bacterial liquid is turbid or there is a large amount of bacterial growth at the bottom, it can be used.
Growth conditions	37 ℃; 24-48 hours; 5% CO2+95% air;
Storage conditions	2-8 ℃
Safety level	1
morphology	For Gram positive bacteria, the MRS liquid culture medium is turbid, with a large amount of sediment at the bottom of the test tube. The growth of the Agar plate strain is weak, and it is recommended to use liquid culture
Sharing mode	Public welfare sharing

Lactobacillus leichmannii

Storage conditions: 2~8 ℃

No. 289804

Product format: freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1 , handle in ultra-clean 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 ℃,5% CO₂,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 cell recovery are reported as follows:

project	test results
viability	Good viability, in 24-48h the liquid medium is turbid, and strain grow slowly.
colony morphology: (above)	Lactobacillus lementi, Gram-positive bacilli, MRS liquid culture bacterial fluid is 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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