Mach1-T1 Escherichia coli|357915 |BNCC

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Escherichia coli Mach1-T1

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	37 ℃; 18-24h; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	Pure
Sharing mode	Public welfare sharing

Escherichia coli Mach1-T1

Storage conditions : 2~8 ℃

No. : 357915

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. Activation

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. Mach1-T1 a single colony is selected from the E. coli 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 should be between 0.4~0.5, and cell number must ﹤ 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.

7.200 μl with ice-cold 0.1mol/l CaCl2 suspension cells.

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

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