Rough chain fungus|336675 |BNCC

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Neurospora crassa

Subculture procedure	① Prepare one test tube containing 5-10 mL of liquid culture medium and two 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 Transfer 0.5mL of liquid culture medium into a freeze-drying tube, dissolve it thoroughly, and then transfer it back into the liquid test tube, mixing well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all liquid test tubes and plates under the above cultivation conditions, and the bacterial strains can be used once they grow.
Growth conditions	28 ℃; 5-7 days; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	Filamentous fungi have obvious colonies on comprehensive PDA medium, and their hyphae spread and grow vigorously. In the early stage, the hyphae are white and dense, and in the later stage, they produce orange yellow, sparse and long hyphae, and orange yellow spores
Sharing mode	Public welfare sharing

Neurospora crassa

Storage conditions: 2~8 ℃

No. 336675

Product format: freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1 , handle in ultra-clean 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 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 lose of viability.

Growth conditions :25-28 ℃, aerobic, integrated PDA,5-7 days. Comprehensive PDA: potato boiling solution 1.0L, glucose 20.0g,KH2PO4 3.0g,MgSO4·7H2O 1.5g, vitamin B1 trace, agar 20.0g,pH 6.0±0.2. Sterilization at 121 ℃ for 15min. Potato boiling liquid: weigh 200g of peeled potato pieces, boil in boiling water for 30min, and collect the filtrate to a constant volume of 1.0L.

Recovery steps:

(1)Prepare the two plates;

(2)Open 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 result
viability	good viability, in 5-7 days strain layer is obvious and colony typical
colony morphology: (above)	filamentous fungi have obvious colonies on the integrated PDA medium, and the hyphae are vigorous and spread. The initial hyphae are white and dense, and the later hyphae are orange, sparse and long, and orange spores are produced.
conclusion	good viability, no abnormal colony morphology, qualified

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