pCMV-3×HA-Myc-MCS-Neo - Basic Information

Recommended Medium	Yeast Extract: 5.0 g, Peptone: 10.0 g, NaCl: 10.0 g, pH: 7.0 (25℃)
Growth Conditions	37℃； 18-24 h； Aerobic
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20 μL of sterile water to the plasmid dry powder and allow it to stand at room temperature for 1 min; ② Mixing: Mix 200 μL of competent cells with 5-10 μL of plasmid DNA and place on ice for 30 min; ③ Heat-shock transformation: Place at 42℃ for 90 s; ④ Membrane pore contraction: Place on ice for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube and culture at 37℃ on a shaker for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of the recovered cells onto an LB plate containing the corresponding antibiotic. Leave the plate 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 a single colony and inoculate it into liquid Recommended Medium with the corresponding antibiotic. Culture with shaking for 12-16 h, then extract the plasmid as required for the experiment.
Reference Application	pCMV-3×HA-Myc-MCS-Neo is a plasmid vector designed for protein expression in mammalian cells. It utilizes the CMV promoter to drive the expression of the target gene fused with both 3×HA and Myc dual tags, and carries a neomycin (Neo/G418) resistance gene for stable cell line selection.
Sharing Policy	Public welfare sharing