pCMV-MCS1-EGFP-CAG-MCS2-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 dry powder for dissolution, 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 Selection culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding antibiotic, leave the plates upright for 30min (the agar surface must be dried), 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-MCS1-EGFP-CAG-MCS2-Neo is a dual-promoter protein expression vector for mammalian cells, utilizing two strong promoters, CMV and CAG, to drive the expression of the gene of interest and the EGFP reporter gene at two multiple cloning sites (MCS), respectively, and enabling stable cell line selection through the neomycin (Neo/G418) resistance gene.
Sharing Method	Public Welfare Sharing