Bacillus subtilis subsp. spizizenii Nakamura et al.|BNCC337945 |BNCC

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Bacillus subtilis subsp. spizizenii Nakamura et al.

Culture medium	Nutritional gravy medium (NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O to culture Bacillus is beneficial to produce spores.
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 the flame, quickly drop sterile water to creak it, and 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	37 ℃;18-24 hours; Aerobic
Storage conditions	2-8 ℃
morphology	G + bacillus size: 1-2mm shape: round edge: irregular transparency: opaque color: off-white uplift: flat surface: rough texture: dry
Sharing mode	Public welfare sharing

Bacillus subtilis

Storage conditions: -20 ℃

Validity period: 24 months

No.: 337945

Product format: freeze dried, 200ul

Biosafety level: 1 , handle in ultra-clean table or safety cabinet

Use: pharmacopoeia, GB 4789, control quality control strain

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. The bacterial tablets shall be used up once and shall not be retained once the penicillin bottled is opened. Keep it at -20℃ for storage if it is not used. 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.

Medium BNCC Medium 1: Nutrient gravy medium Beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 20.0g (liquid medium not included), distilled water 1.0L, pH 7.0. Sterilize at 121°C for 15min. [Note] Adding 5mg MnSO4·H2O when culturing Bacillus is beneficial to produce spores.

Growth conditions Temperature: 37 ℃ Gas environment: aerobic

Recovery steps:

(1)Prepare a sterile test tube of 5-10ml liquid medium and 2 pieces of 90mm plates;

(2)Sterilizing of the penicillin-bottle, under aseptic conditions remove the aluminium cap and rubber plug, draw 0.5ml of liquid culture medium into the vial, aspirate to dissolve the bacterial tablets.

(3)Fully dissolved, transfer the solution to the liquid test tube, mix evenly;

(4)inoculate a plate with 0.2ml of the solution, distribute it well in 200ml/plate;

(5)Put all the plates and test tubes under the above culture conditions for cultivation, the plates are not allowed to seal by wrapping. The culture solution is obviously turbid after 18-24 hours, and this indicates the bacterial grows well.