Lactobacillus delbrueckii subsp. bulgaricus|BNCC187939 |BNCC

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Lactobacillus delbrueckii subsp. bulgaricus

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	30 ℃;24-48h; Anaerobic;
Storage conditions	2-8 ℃
morphology	G +, the thallus is long rod-shaped, the ends are blunt and round, arranged in a chain, without spores, and with different stained grains. The colony is round, with rough surface, protrusion, neat edges, light yellow, opaque.
application	Produces yogurt.
Sharing mode	Public welfare sharing

Lactobacillus delbrueckii subsp. bulgaricus

Storage conditions: 2~8 ℃

No. 187939

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 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: 30℃, anaerobic, MRS medium, 24-48h. 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 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.04g , Tween 80 1.0g, agar 20.0g (liquid medium not included), distilled water 1.0L. 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 liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology:	Size: 1-2mm Shape: Irregular Edge: Irregular Transparency: Opaque color: off-white uplift: flat surface: bright and smooth texture: wet and easy
Conclusion:	good viability, no abnormal colony morphology, qualified

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