BL21 (AI) Escherichia coli|356366 |BNCC

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Escherichia coli BL21(AI)

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 two plates, each Agar plate containing about 200 μ L, 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	37 ℃; 18-24h; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	Pure
Sharing mode	Public welfare sharing

Escherichia coli BL21(AI)

Storage conditions :-20℃

No. : 356366

Product format :freeze dried, 200ul

Validity : 5 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 -20 ℃ after receiving it. If any abnormal situation such as damage is found on the same day, please contact customer service within 24 hours. The Xilin bottle shall be used up once after opening 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 ℃

Environment: aerobic

Usage:

Recovery A.

1. Prepare 2 LB plates;

2. under aseptic environment, 0.5mL of liquid culture medium is injected into the freeze-dried tube to fully dissolve the freeze-dried powder; 3. Scribe the suspension in a flat plate, and grow a single colony in 2 flat plates for 18-24 hours;

B. competent preparation (for reference)

1. a single colony was picked from escherichia coli BL21(AI) 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, and culture at 37 ℃ for 2~3h,A600 0.4~0.5 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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