pLV3-MSCV-3×FLAG-MCS-EF1a-CopGFP-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 for dissolution, let stand at room temperature for 1min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, 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, culture at 37℃ on a shaker for 1h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with corresponding resistance, incubate plates upright for 30min (the agar surface must be dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with corresponding resistance, culture with shaking for 12-16h, and extract plasmids according to experimental needs.
Reference Application	pLV3-MSCV-3×FLAG-MCS-EF1a-CopGFP-Puro is a lentiviral expression vector that uses the MSCV long terminal repeat (LTR) to drive expression of the target gene in the multiple cloning site (MCS), fusing a 3×FLAG tag to the N-terminus of the target protein for easy detection.
Sharing Method	Public Welfare Sharing