pBV220|354578 |BNCC

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pBV220

Growth conditions	30°C; 18-24h; aerobic
Subculture procedure	①Dissolution: Add 20 μL sterile water to the plasmid powder for dissolution, let stand at Room temperature for 1 min;② Mixing: Add 200μl competent cells to 5-10μl plasmid DNA, mix thoroughly, and incubate on ice for 30 min; ③ Heat shock transformation: Incubate at 42°C for 90 s; ④ Shrink membrane pores: Ice bath for 2 min;⑤ Repair culture: Add 800 μl liquid medium to each tube; incubate on a 37°C shaker (150 rpm) for 1 hour;⑥ Screening Culture: Spread an appropriate volume (100 μl) of the resuspended cells onto LB plates with the corresponding antibiotic. Incubate plates upright for 30 minutes (ensure agar surface is completely dry), then invert and incubate for 12-16 hours until colonies appear.⑦ Extraction: Transfer a single colony to the corresponding antibiotic-containing LB liquid medium. Incubate with shaking for 12–16 hours. Extract the plasmid as required for the experiment.
Storage conditions	2-8°C
Sharing mode	Public welfare sharing