Hooked Agrobacterium|134955 |BNCC

Essential Information Certificate Related Products

Agrobacterium rubi

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	30 ℃; 18-24h; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The bacterium forms small (0.5–1 mm), circular, opaque, convex colonies with entire margins, a smooth, light red surface, and a moist texture. Microscopic examination reveals Gram‑negative (red) bacilli.
Sharing mode	Public welfare sharing

Rhizobium vitis

Storage conditions: 2~8 ℃

No. 134955

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: 30℃, aerobic, 24h, soybean rhizobia culture medium: mannitol 10-20 g, potassium dihydrogen phosphate (KH2PO4) 0.5 g, sodium chloride (NaCl) 0.2 g, magnesium sulfate (MgSO4 7H2O) 0.2 g, Calcium sulfate (CaSO4·2H2O) 0.1 g, calcium carbonate (CaCO3) 1.0 g, yeast liquid (10:100) 100 ml, agar 18-20 g, clear tap water 900 ml. 121℃, 15min.

Recovery steps:

(1)Prepare 1-2 pieces of above mentioned plates;

(2)Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip in a flame, quickly drop sterile water to creak it, then break it with forceps;

(3)Draw 0.5ml of sterile water into the freeze dried ampoule, make it fully dissolved, and distribute the solution to the plates well in 200ul/plate;

(4)Put the plates under the above culture conditions for cultivation for18-24 hours.

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

Item	test results
viability	good viability，in 20 hours,liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology:	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Translucent Color: off-white uplift: middle convex surface: bright and smooth texture: moist and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified

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