pCMV-EGFP-MCS2-3×Myc-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 h; Aerobic
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20 μL sterile water to the plasmid dry powder 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°C 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 at 37°C with shaking 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, keep plates upright for 30 min (ensure the agar surface is completely 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 and culture with shaking for 12-16 h, then extract plasmid as needed for the experiment.
Intended Use	pCMV-EGFP-MCS2-3×Myc-Neo is a fluorescent fusion vector for mammalian cell protein expression. It uses the CMV promoter to drive fusion expression of EGFP with the target gene, carries a 3×Myc tag at the C-terminus of the target gene, and also contains the neomycin (Neo/G418) resistance gene for stable cell line selection.
Sharing Method	Public welfare sharing