Fusobacterium nucleatum subspecies|280188 |BNCC

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Fusobacterium nucleatum subsp.nucleatum Knorr

Subculture procedure	Prepare 1-2 of the above plates (placed in an anaerobic environment for 24 hours of deoxygenation in advance) with typical bacterial colonies; ② Open the 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 or liquid culture medium (placed in an anaerobic environment for 24 hours of deoxygenation in advance) 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/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 ℃; 3-5 days; anaerobic
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony diameter is 0.5-1mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, smooth and bright on the surface, moist texture, easy to pick up, G - (red), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Fusobacterium nucleatum subsp.

Storage conditions: 2~8 ℃

No. 280188

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level: 2, handle in 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°C, anaerobic, thioglycolate broth, 3-5 days. Thioglycolate liquid medium: caseptone (trypsin hydrolysis) 15.0g, yeast extract 5.0g, glucose 5.0g, sodium thioglycolate 0.5g, L-cystine 0.5g, sodium chloride 2.5g, resate Green 0.001g, agar 0.75g, distilled water 1.0L, pH 7.1±0.2. Sterilize at 121℃ for 15min.

Recovery steps:

(1)Prepare a φ18mm test tube of 10ml liquid medium; and two agar plates

(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 3 days,liquid medium become turbid
colony morphology: (above)	Fusobacterium nucleated subspecies, Gram-negative bacteria, Clostridium, thioglycolate liquid medium culture bacteria liquid turbid, it is recommended to use liquid culture.
Conclusion	good viability, no abnormal colony morphology, qualified

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