pCMV-MCS-V5-Neo-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 lyophilized powder to dissolve, and let stand at room temperature for 1min； ② Mixing: Take 200μl competent cells and mix 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 Selection culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding antibiotic, keep the plates upright for 30min (the agar surface must be thoroughly dry), then invert and culture for 12-16h until colonies appear； ⑦ Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid according to experimental needs.
Reference Application	pCMV-MCS-V5-Neo is a plasmid vector used for protein expression in mammalian cells. It utilizes the CMV promoter to drive the expression of the target gene fused with the V5 tag, and carries a neomycin (Neo/G418) resistance gene for stable cell line selection.
Sharing Method	Public Welfare Sharing