pECMV-Nox4-m-FLAG - Basic Information

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