AGL1 Agrobacterium|356354 |BNCC

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Agrobacterium tumefaciens AGL1

Subculture procedure	① Prepare 1-2 of the above 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 Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	28 ℃; 48-72h; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, smooth on the surface, moist texture, G - (red), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Agrobacterium tumefaciens AGL1

Storage conditions : 2~8 ℃

No. : 356354

Product format :freeze dried, 200ul

Validity : 6 years

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

Uses: competent cells, plasmids, genetic engineering

Notes: this product is transported at room temperature. please store it at 2-8 ℃ after receiving it. If any abnormal situation such as damage is found on the same day, please contact customer service within 24 hours. The freeze-dried powder shall be used up once after being opened and shall not be retained. Please operate in strict accordance with this instruction, otherwise the replacement service will not be provided if the strain is abnormal and inactivated.

Medium :LB agar medium yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, agar 15.0g, distilled water 1.0 L,pH 7.0. Sterilization at 121 ℃ for 15min.

Growth conditions: Temperature: 37 ℃ Gas environment: aerobic

How to use

A. Recovery 1. Prepare 2 LB plates;

2. In a sterile environment, insert 0.5mL of liquid culture medium into a lyophilized tube to fully dissolve the lyophilized powder;

3. Scribe the suspension on the plate, 2 plates, 18-24h to grow a single colony;

B. competent preparation (for reference)

1. a single colony was picked from agrobacterium AGL1 plate and connected to a test tube of 3ml LB liquid medium, and cultured overnight at 37 ℃.

2. take 1ml of bacterial solution and transfer it to a conical flask containing 50ml LB liquid culture medium. shake culture at 37 ℃ for 2~3h,A600 should be between 0.4~0.5, and the number of cells must be <108CFU/ml.

3. transfer 1ml of bacterial liquid to a 1.5 ml centrifuge tube with a pipette gun and place it on ice for 10min.

4. Centrifugation for 10min,4000r /min.

5. pour out the culture solution and use cold 0.1mol/l CaCl2 200 μL suspension precipitation, immediately placed on ice for 30min.

6. Low temperature centrifugation for 10min,4000r /min, cells were recovered.

7. 200µl with ice-cold 0.1mol/l CaCl2 200 μLsuspension cells.

8. subpackage cells, each 200ul, this cell is competent cells.

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