pLV3-CMV-3×FLAG-MCS-EF1a-mCherry-Puro - 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 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 resistance. Leave the plates upright for 30min (ensure the agar surface is completely dry first), then invert and culture for 12-16h until colonies appear； ⑦ Extraction: Pick a single monoclonal colony into LB liquid Recommended Medium with the corresponding antibiotic resistance, culture with shaking for 12-16h, and extract the plasmid as needed for the experiment.
Intended Use	pLV3-CMV-3×FLAG-MCS-EF1a-mCherry-Puro is a dual-expression cassette lentiviral vector for stable expression in mammalian cells. It uses the CMV promoter to drive the fusion expression of the target gene with a 3×FLAG tag, while the EF1a promoter independently expresses mCherry red fluorescent protein and puromycin resistance for dual selection and tracking.
Sharing Method	Public welfare sharing