Escherichia coli Rosetta(DE3) Strain|BNCC341754 |BNCC

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Escherichia coli Rosetta(DE3) Strain

Culture medium	LB +10 μg/ml chloramphenicol: LB +10 μg/ml chloramphenicol: yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. After sterilization, chloramphenicol was added at a concentration of 10 μg/ml when the culture medium was cooled to 40-50 ℃.
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 ℃
morphology	Colony shape: size: 1-2mm shape: round edge: neat transparency: opaque color: off-white uplift: flat surface: smooth gray texture: moist and easy to stir up
application	for protein expression
Sharing mode	Public welfare sharing

Rosetta(DE3) Strain Rosetta(DE3) Strain

Storage conditions: 2~8 ℃

No. 341754

Product format: freeze dried, 200ul

Validity : 6 years

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

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions:37 ℃, aerobic, LB agar medium, 18-24h. LB agar medium: yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, agar 15.0g (not included in liquid medium), distilled water 1.0 L,pH 7.0. Sterilization at 121 ℃ for 15min. Chloramphenicol was added when the culture medium was cooled to 50 ℃ to make its working concentration 50ug/ml chloramphenicol.

Recovery steps:
(1) Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;
(2) Open the ampoule in the biosafety 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.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability，in 24 hours liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	size: 1-2mm ;Shape: round ;Edge: neat; Transparency: opaque color: off-white ; Protuberance: flat ; Surface: smooth and gray ; Texture: moist and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified