pCMV-Nluc - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	37℃； 18-24h； Aerobic
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1min； ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, and 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 on LB plates with the corresponding resistance. Leave the plates upright for 30min (the agar surface must be thoroughly dry first), then invert and culture for 12-16h until colonies appear； ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding resistance, culture with shaking for 12-16h, and extract the plasmid according to experimental needs.
Reference Use	pCMV-Nluc is a NanoLuc luciferase reporter plasmid for mammalian cells, driven by the strong CMV promoter, and uses ampicillin as the prokaryotic resistance selection marker.
Sharing Method	Public welfare sharing

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