Mycobacterium aurantium|BNCC360708 |BNCC

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Mycobacterium aurantium

Culture medium	7H10 agar medium (English name: 7H10): 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, absorb 5ml of glycerol and add 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 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 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	30°C;3-4 days; Aerobic
Storage conditions	2-8 ℃
morphology	The diameter of the colony is 1-2mm, the shape is irregular, the edge is irregular, opaque, the front is off-white, the back is off-white, the shape is flat, the surface is rough, the texture is dry, it is easy to stir up, the acid-fast staining is positive (red), bacilli, Purity: pure
Sharing mode	Public welfare sharing

Mycobacterium marinum

Strain number: 360708

Batch number: 220531

Biosafety level: 2, handle in safety cabinet

Provide form: Freeze-dried beveled bacterial liquid plate

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. The freeze-dried powder is stored at 2~8℃, and the validity period is 6 years; the slant surface is stored at 2~8℃, and the validity period is 1~3 months; the plate and bacterial liquid are stored at 4℃, and the validity period is 7 days. 2 years. Please strictly follow these instructions for activation and preservation, otherwise, the reissue service will not be provided due to abnormal bacteria, inactivation, etc.

Medium: 7H10 agar medium (English name: 7H10): ammonium sulfate 0.5g, potassium dihydrogen phosphate 1.5g, disodium hydrogen phosphate 1.5g, sodium citrate 0.4g, magnesium sulfate 0.025g, calcium chloride 0.5mg, zinc sulfate 0.001 g, copper sulfate 0.001g, sodium L-glutamate 0.5g, ferric ammonium citrate 0.04g, pyridoxine hydrochloride 0.001g, biotin 0.5mg, malachite green 0.25mg, agar 15.0g, and draw 5ml of glycerol, Add 900ml of distilled water, heat and stir to dissolve, boil for 1 minute, pH 6.4-6.8. Autoclave at 121°C for 10 minutes. When cooled to 50-55°C, add 100ml of filter-sterilized OADC additive, mix well, and pour into a plate or test tube.

Growth conditions: 30°C;3-4 days; aerobic

Recovery steps:

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

(2)Open the ampoule in the 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 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.

Viability: good viability, 3-5 days, with obvious bacterial layer on the plate; colony is typical on the streaked plate

Morphological characteristics: Colony diameter is 1-2mm, irregular shape, irregular edge, opaque, front gray-white, reverse gray-white, flat shape, rough surface, dry texture, easy to provoke, acid-fast staining positive (red), bacilli, purity: pure

Quality inspection map:

Conclusion:good viability， no abnormal colony morphology, qualified

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