Escherichia coli DH5α(pACYC184)|BNCC353850 |BNCC

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Escherichia coli DH5α(pACYC184)

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 1 sterile test tube of 5~10mL of liquid medium and 2 pieces of 90mm plates; 2. sterilizing of the ampoule, 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 the ampoule, fully dissolved, transfer the solution to the liquid test tube, mix evenly; 4. inoculate a plate with 0.2ml of the solution, distribute it well in 200ml/plate; 5. Put all the plates and test tubes under the above culture conditions for cultivation, the plates are not allowed to seal by wrapping. The culture solution is obviously turbid after 18-24 hours, and this indicates the bacterial grows well;
Growth conditions	37 ℃;18-24h; Aerobic;
Storage conditions	2-8 ℃
application	Scientific research
Sharing mode	Public welfare sharing

Escherichia coli DH5α(pACYC184)

No.: 353850

Storage conditions: 2~8 ℃

Product format: freeze dried, 200ul

Validity period: 6 years

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

Notes: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. The bacterial tablets shall be used up once and shall not be retained once the penicillin bottled is opened. Keep it at -20℃ for storage if it is not used. 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.

Host bacteria: 353719 DH5α;.

Plasmid：353817 pACYC184 (containing chloramphenicol resistance gene).

Medium:LB resistant agar medium

yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, agar 15.0g (without liquid medium), distilled water 1.0 L,pH 7.0. Sterilization at 121 ℃ for 15min. After sterilization, cooling and cooling, chloramphenicol is added to make the final content of chloramphenicol in the culture medium 50μg/mL.

Growth conditions:37 ℃ aerobic 18-24h.

Recovery steps:
(1)Prepare a sterile test tube of 5-10ml liquid medium and 2 pieces of 90mm plates;
(2)Sterilizing of the penicillin-bottle, under aseptic conditions remove the aluminium cap and rubber plug, draw 0.5ml of liquid culture medium into the vial, aspirate to dissolve the bacterial tablets.
(3)Fully dissolved, transfer the solution to the liquid test tube, mix evenly;
(4)Inoculate a plate with 0.2ml of the solution, distribute it well in 200ml/plate;
(5)Put all the plates and test tubes under the above culture conditions for cultivation, the plates are not allowed to seal by wrapping. The culture solution is obviously turbid after 18-24 hours, and this indicates the bacterial grows well.
(6) If it does not grow in 18-24 hours, culture is continued until 72 hours is reached. And the strains can be used when they grow

plasmid purification:

1. Pick a single colony and inoculate it into 50mL of liquid LB resistance medium, and culture at 37 ℃ overnight.

2. According to the experimental requirements, absorb appropriate amount of bacterial liquid for plasmid extraction.

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