JM110 Escherichia coli|357910 |BNCC

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Escherichia coli JM110

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	The colony diameter is 1-2mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, smooth and bright on the surface, moist texture, easy to pick up, G - (red), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Escherichia coli JM110

Storage conditions : 2~8 ℃

No. : 357910

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

* Note : 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. under sterile conditions, draw 0.5mL of liquid culture medium into a lyophilized tube to fully dissolve the freeze dried pellets;
3.inoculate two plates with the liquid suspension, and streak the two plates, in 18-24h a single colony grow ;

B. Competent preparation (for reference)
1. inoculate a test tube of 3ml LB liquid medium with a single colony from JM110(DE3) E. coli plate , and put the test tube on a shaker at 37 ℃ overnight.
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 and place it on ice for 10min.
4. Centrifugation for 10min,4000r /min.
5. suck out all the supernatant and add 200ul of ice-cold CaCl ( 0.1mol/l), then immediately place on ice for 30min.
6. low temperature centrifugation for 10min,4000r /min to collect cells.
7. suspend the cells with 200ul of ice-cold CaCl ( 0.1mol/l).
8. sub-package cells, each 200ul, this is competent cells.

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