pCMV-MCS-CFP-Neo - Basic Information

Recommended Medium	Yeast extract: 5.0 g, Peptone: 10.0 g, NaCl: 10.0 g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24 hours; Aerobic;
Storage Conditions	2-8°C
Procedure	① Dissolve: Add 20 μL sterile water to the plasmid dry powder for dissolution, and let stand at room temperature for 1 min; ② Mix: Take 200 μL competent cells and mix with 5-10 μL plasmid DNA, and place on ice for 30 min; ③ Heat-shock introduction: Let stand at 42°C for 90 s; ④ Contract membrane pores: Ice bath for 2 min; ⑤ Recovery culture: Add 800 μL liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1 h (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 30 min (the agar surface must be dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single colonies into liquid LB Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid according to experimental needs.
Reference Application	pCMV-MCS-CFP-Neo is a plasmid vector used for mammalian cell expression. It uses the CMV promoter to drive fusion expression of the target gene with cyan fluorescent protein (CFP), and carries a neomycin (Neo/G418) resistance gene for stable cell line selection.
Sharing Method	Public welfare sharing