Mycobacterium intracellular|BNCC359486 |BNCC

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Mycobacterium intracellular

Culture medium	7H10 agar medium (7H10): 0.5g of ammonium sulfate, 1.5g of potassium dihydrogen phosphate, 1.5g of disodium hydrogen phosphate, 0.4g of sodium citrate, 0.025g of magnesium sulfate, 0.5mg of calcium chloride, 0.001g of zinc sulfate, 0.001g of copper sulfate, 0.5g of sodium L-glutamate, 0.04g of ammonium citrate, 0.001g of pyridoxine hydrochloride, 0.5mg of biotin, 0.25mg of malachite green, 15.0g of agar, 5ml of glycerol and 900ml of 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.
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 ℃;3-5 days; Aerobic;
Storage conditions	2-8 ℃
morphology	Colony diameter of 1-2mm, round, irregular edges, opaque, yellow on the front, convex in the middle, smooth surface, moist texture, G + (blue-purple), bacilli, purity: pure
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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