Escherichia coli BL21(DE3)(pET-32a(+)-Proteinase

Essential Information Certificate Related Products

Escherichia coli BL21(DE3)(pET-32a(+)-Proteinase_3CL-pro)

Culture medium	LB + 50mcg/ml ampicillin: yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. After sterilization, ampicillin was added when the culture medium was cooled to 40-50 ℃ with a concentration of 50mcg/ml.
Subculture procedure	1. Prepare 1 sterile test tube of 5~10mL of 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, blow 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.
Growth conditions	37 ℃;18-24h; Aerobic;
Storage conditions	-20 ℃
application	pET-32a(+)-Proteinase_3CL-pro plasmid extraction. Gene species: SARS-CoV-2
Sharing mode	Public welfare sharing

Escherichia coli BL21(DE3) ( PET-32a(+)-Proteinase_3CL-pro )

No. 354684

Product format: tablets in penicillin bottle

Storage conditions: -20 ℃

Biosafety level: level 1

Validity: 5 years

Purpose: Plasmid extraction

Gene species: SARS-CoV-2 *

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.

Medium:LB resistant 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. After sterilization, add ampicillin sodium after cooling and cooling, so that the final content of ampicillin sodium in the culture medium is 50 ug/mL.

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, blow 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.

Growth condition temperature:37 ℃

Gas environment: Aerobic

Time:18-24h