Stbl3 (containing pLV3-U6-sgRNA-U6-sgRNA-Cas9-EGFP-Puro plasmid) E. coli — Basic Information

Recommended Medium	Yeast extract: 5.0 g, Peptone: 10.0 g, NaCl: 10.0 g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24 h; Aerobic
Storage Conditions	-80°C
Procedure	① Thaw the cryovial in a water bath (mold: 30°C, bacteria: 37°C) and shake rapidly to dissolve; ② Wipe the outside of the cryovial with an alcohol cotton pad for disinfection, then transfer it to a biosafety cabinet for handling; ③ Unscrew the cap, aspirate all of the dissolved suspension, and spread it onto 2 plates with the corresponding antibiotic, approximately 200 μL per plate; ④ Incubate the antibiotic plates under the specified conditions above for 18-24 h, then observe (do not seal or wrap the plates); ⑤ Pick a single colony and inoculate it into 50 mL of liquid LB with the corresponding antibiotic in the Recommended Medium, and culture overnight at 37°C on a shaker; ⑥ According to experimental needs, take an appropriate amount of the bacterial culture for plasmid extraction.
Sharing Method	Public welfare sharing