Javanese cordyceps|228354 |BNCC

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Cordyceps javanica

Subculture procedure	① Dissolve the frozen tube in a water bath (fungi at 30 ℃, bacteria at 37 ℃) and shake quickly for dissolution; ② Wipe the outer wall of the cryovial with an alcohol swab for disinfection, and then transfer it to a safe cabinet for operation; ③ Unscrew the tube cap, aspirate all the dissolved solution, and transfer it into 1-2 agar plates (each plate contains approximately 200 μL, and for anaerobic bacteria cultivation, the culture medium needs to be placed in an anaerobic environment for 24 hours to remove oxygen); ④ After spreading evenly, transfer to the above cultivation conditions for cultivation.
Growth conditions	28 ℃, 3-5 days, aerobic
Storage conditions	Liquid nitrogen
Safety level	1
morphology	Small filamentous fungi forming distinct colonies on PDA medium. Colonies appear white with dense and vigorous mycelial growth, and are asporogenous. The reverse side of the medium is pale yellow.
Sharing mode	Public welfare sharing

Isaria tenuipes

BNCC No.： 228354

Storage conditions: 2~8 ℃

Product format：agar slant in 14mm test tube

Validity period: growing culture, in 30 dyas

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 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 ℃, aerobic, integrated PDA,3-5 days. Comprehensive PDA: potato boiling solution 1.0L, glucose 20.0g,KH₂PO₄ 3.0g,MgSO_4·7H₂O 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 1-2 pieces of PDA plates;

(2)Sterilize the surface of test tube for the agar slant, open it in the biosafety cabinet;

(3)Cut into the pieces with aseptic inoculation hoe and inoculation shovel (see attached page). the size of square pieces is 0.5 × 0.5cm2;

(4)Lay flat the small pieces to the center of the agar plate;

(5)Put the plates under the above culture conditions, 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 3-5 days strain layer is obvious
colony morphology	small filamentous fungi with obvious colonies on comprehensive PDA medium, White, dense and vigorous hyphae, spreading to the edge of the plate and not producing spores.
conclusion	good viability, no abnormal colony morphology, qualified

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