Pichia pastoris|BNCC225716 |BNCC

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Pichia pastoris

Culture medium	YM medium (YM): yeast extract 3.0g, malt extract 3.0g, glucose 10.0g, peptone 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 6.2±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare 1-2YM 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, inoculate a plate with the solution at 200ul/plate; (4) Put the plate under the above culture conditions for18-24 hours.
Growth conditions	30 ℃,24-48h, aerobic
Storage conditions	2-8 ℃
morphology	Size: General shape: round edge: neat edge transparency: opaque color: milky white uplift: raised surface: smooth and bright texture: moist
application	Efficient expression, secretion and production of lipase
Sharing mode	Public welfare sharing

Pichia pastoris

Storage conditions: 2~8 ℃

BNCC No.：225716

Product format： freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear table or 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 slant and suspension liquid 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:28-30 ℃, aerobic, YM medium. YM medium: yeast extract, 3.0g malt extract, 3.0g glucose, 10.0g protein block, 5.0g agar, 20.0g distilled water 1000 ml ,pH 6.2 +/- 0.2,121 ℃,15min.

Recovery steps:

(1) Prepare 1-2 YM 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) Put the agar plates under the above culture conditions for cultivation 24-48 hours.

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

item	test result
viability	good viability, in 40h YM plate colony is obvious
colony morphology: (above)	size: general shape: round edge: neat edge transparency: opaque Color: milky white uplift: raised surface: smooth and bright texture: moist
Conclusion	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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