pMV261 - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	LB + antibiotic, 37℃
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder for dissolution, and let stand at room temperature for 1min; ② Mixing: Take 200μl competent cells and mix with 5-10μl plasmid DNA, then place on ice for 30min; ③ Heat-shock introduction: Let stand at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture on a shaker at 37℃ for 1h (150 r/min); ⑥ Screening culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding antibiotic, leave plates upright for 30min (the agar surface must be completely dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick a single monoclonal colony into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid according to experimental needs.
Sharing Method	Public welfare sharing