Lactococcus lactis subsp. lactis|BNCC185907 |BNCC

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Lactococcus lactis subsp. lactis

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 ℃;24-48h; Anaerobic;
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with irregular edges, opaque, white on the front, golden yellow on the back, convex in the middle, smooth and bright surface, moist texture, easy to stir up, no pigment, G + (blue-purple), cocci, purity: pure
application	Used in Swiss cheese production to inhibit the production of Clostridium species. Nisin is produced.
Sharing mode	Public welfare sharing

Lactococcus lactis subsp

No. 185907

Storage conditions: 2~8 ℃

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:37 ℃, anaerobic, 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 0.05g, Tween 80 1.0g,L-cysteine 0.5g, agar 20.0g (not included in liquid medium), distilled water 1.0L. pH 6.2±0.2. 121 ℃,15min sterilization

Recovery steps:

(1)Prepare sterile test tube of 5-10ml liquid medium and 2 plates; Silica gel plug is used for liquid test tube culture medium to ensure ventilation and facilitate deoxygenation. Please put the liquid medium and plate in an anaerobic environment for 24h before usage.

(2)Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip 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)Put the plates under the above culture conditions for cultivation for 24-48 hours.

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 liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	size: 1-2mm shape: round edge: irregular transparency: opaque color: white uplift: middle raised surface: bright and smooth texture: moist and viscous
conclusion:	good viability, no abnormal colony morphology, qualified