Intracellular mycobacteria|359486 |BNCC

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Mycobacterium intracellulare

Subculture procedure	① Prepare 1-2 of the above plates; 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 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 per 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; aerobic;
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony diameter is 0.5-1mm, circular, with irregular edges, opaque, golden yellow on the front, raised in the middle, smooth on the surface, moist texture, easy to pick up, G+(blue purple), rod-shaped
Sharing mode	Public welfare sharing

Mycobacterium intracellular

Storage conditions : 2~8 ℃

No. : 359486

Product format :freeze dried, 200ul

Validity : 6 years

Biosafety level : 2, handle in 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 plate 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 ℃, aerobic, 7H10 agar medium. For 3-5 days, 7H10 agar medium: 0.5g ammonium sulfate, 1.5g potassium dihydrogen phosphate, 1.5g disodium hydrogen phosphate, 0.4g sodium citrate, 0.025g magnesium sulfate, 0.5mg calcium chloride, 0.001g zinc sulfate, 0.001g copper sulfate, 0.5g L-sodium glutamate, 0.04g ferric ammonium citrate, 0.001g pyridoxine hydrochloride, 0.5mg biotin, 0.25mg malachite green, 15.0g agar, 5ml glycerol and 900ml distilled water, heat, stir and dissolve, boil for 1 minute with a PH value of 6.4-6.8. High pressure sterilization at 121 ℃ for 10 minutes. When cooling 50-55 ℃, add 100ml of OADC additive for filtration and sterilization, mix well, and pour into a plate or test tube.

Note:OADC additive preparation (100ml): 5g of bovine albumin (component v), 0.06ml of oleic acid, 2g of glucose, 0.003g of enzyme and 0.85g of sodium chloride.

Recovery steps:

① Prepare a test tube containing 5~10mL of liquid culture medium and 2 plates;

②Open it in the safety cabinet, burn the top with an alcohol lamp, quickly drop sterile water to break it, and then break it with tweezers;

③ suck 0.5mL of liquid culture medium into freeze-dried tube, fully dissolve and return to liquid test tube again, and mix well;

④ Absorb 0.2mL of bacterial suspension into the plate, coat evenly, and repeat twice to obtain two plates;

⑤ Place all the liquid test tubes and plates under the above culture conditions, and the strains can be used when they grow out.

Recovery record: activation is carried out according to activation requirements, and single colony is observed by scribing. the record results are as follows:

Item	test results
viability	good viability, in 3days,strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Opacity color: yellow uplift: middle raised surface: smooth texture: wet
Conclusion	good viability, no abnormal colony morphology, qualified

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