|Culture medium||LB + 50mcg/ml kanamycin: yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. After sterilization, kanamycin was added when the culture medium was cooled to 40-50 ℃ at a concentration of 50mcg/ml.|
|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 Stbl3(pCAMBIA1303-TrpC-Hygro-gpdA-GFP)
Storage conditions: 2~8 ℃
Product format: freeze dried,200ul
Validity period: 6 years
Biosafety level: 1,handle in ultra-clear 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. 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:BNCC353822 Stbl3
Plasmid:BNCC357700 pCAMBIA1303-TrpC-Hygro-gpdA-GFP (containing kanamycin resistance gene).
Medium:LB resistant agar medium Yeast extract 5.0 g, peptone 10.0 g, NaCl 10.0 g, agar 15.0 g (not included in liquid medium), distilled water 1.0 L, pH 7.0. Sterilize at 121 °C for 15 min. After sterilization, after cooling down, kanamycin was added to make the final content of kanamycin in the medium to be 50 μg/mL.
Growth conditions:37 ℃ aerobic 18-24h.
① Prepare a sterile test tube of 5-10ml liquid medium and 2 pieces of 90mm plates;
② Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip 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 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.
1. Pick a single colony and inoculate it into 50mL liquid LB resistance medium, and shake it at 37℃ overnight;
2. According to the experimental requirements, draw an appropriate amount of bacterial liquid for plasmid extraction.