LOADING
Bacterial Preservation: Mix 500 μl of 40% glycerol with 500 μl of bacterial solution (or 300 μl of 50% glycerol with 700 μl of bacterial solution). A total of eight tubes of Amp30E (Kanamycin-resistant strain) were preserved and stored at -80 ℃.
Notes: The operation shall be conducted in a biological safety cabinet, and the glycerol must be sterilized in advance. After transferring the mixture into glycerol storage tubes, mix it thoroughly by pipetting up and down or using a vortex mixer.
Medium Preparation: LB medium was prepared for culturing our chassis strain Escherichia coli (E. coli). Based on the plasmids used (Kanamycin-resistant pSB3K3 and Ampicillin-resistant pSB4A3), we prepared corresponding single-resistant LB solid medium plates, double-resistant LB solid medium plates, and LB liquid medium for shake flask culture of E. coli. The formula of LB medium is included in our Experiments Page.
Sterilization: Sterilize using an autoclave at 121°C for 15 minutes
Notes: When in use, ensure the water level does not exceed the "HIGH" warning line. Regularly drain excess condensed water. When the temperature drops to approximately 70 ℃, the sterilization program can be terminated, and the sterilized medium can be taken out.