Lactococcus lactis subsp. lactis|185907 |BNCC

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Lactococcus lactis subsp. lactis

Subculture procedure	① Prepare 1-2 plates as described above, along with sterile water (pre‑equilibrated in an anaerobic environment for 24 h to remove oxygen).② In a biosafety cabinet, open the vial and briefly flame the top with an alcohol lamp, then immediately add sterile water to break the seal and crush the vial using sterile forceps.③ Aspirate 0.5 mL of the pre‑treated sterile water into the freeze‑dried vial, thoroughly dissolve the bacterial powder, then transfer 200 μL of the suspension onto each Agar plate and spread evenly.④ Incubate the plates under the specified culture conditions. The strain is ready for use once growth appears.
Growth conditions	37 ℃; 24-48 hours; anaerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colonies measure 1-2 mm in diameter, are circular with irregular margins, and are opaque. They appear white on the surface (obverse) and golden yellow on the reverse side, with a raised center. The colonies are smooth, glossy, and moist in texture, making them easy to lift. No pigment is produced. The organisms are Gram-positive (G⁺, appearing blue-violet).
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

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