Mycobacterium chelonae|BNCC359488 |BNCC

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Mycobacterium chelonae

Culture medium	7H10 agar medium (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-glutamate sodium, 0.04g ferric ammonium citrate, 0.001g pyridoxine hydrochloride, 0.5mg biotin, 0.25mg malachite green, 15.0g agar, 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	30 ℃;48-72h; Aerobic;
Storage conditions	2-8 ℃
morphology	Round, irregular edges, opaque, white on the front, light color, convex in the middle, smooth and bright surface, moist texture, easy to stir up, no pigment, bacilli, G + (blue-purple), purity: pure
Sharing mode	Public welfare sharing

Mycobacterium chelonae

Storage conditions: 2~8 ℃

No. 359488

Product format: freeze dried,200ul

Validity period: 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:30°C, aerobic, 7H10 agar medium. 48-72h, 7H10 agar medium: 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 0.001g, 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 5ml glycerol was drawn, added 900ml In 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 100 ml of filter-sterilized OADC additive, mix well, and pour into a plate or test tube. Note: OADC additive preparation (100ml): bovine albumin (V component) 5g, oleic acid 0.06ml, glucose 2g, catalase 0.003g, sodium chloride 0.85g.

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.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability. in 72 hours obvious strain layer occurs on the plate; colony is typical
Colony morphology: (pictured above)	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Opaque Color: White Protuberance: Middle Protuberance Surface: Smooth Texture: Wet and Easy to Stir
conclusion: good viability, no abnormal colony morphology, qualified recovery

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