Rosetta(DE3) Escherichia coli|BNCC357922 |BNCC

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Rosetta(DE3) Escherichia coli

Culture medium	Base medium (English name: LB): yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps; (3) draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly; (4) inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates; ⑤ Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Growth conditions	37 ℃;18-24h; Aerobic;
Storage conditions	2-8 ℃
Sharing mode	Public welfare sharing

Escherichia coli Rosetta(DE3)

Storage conditions : 2~8 ℃

No. : 357922

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.Rosetta(DE3) E. coli plate, pick a single colony and connect it to a test tube of 3ml LB liquid medium, and culture at 37 ℃ for overnight.

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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