pLV3-CMV-MCS-EF1a-mTagBFP2-Puro - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24h; Aerobic
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20μL of sterile water to the plasmid lyophilized powder to dissolve, and let stand at room temperature for 1min; ② Mixing: Mix 200μl of competent cells with 5-10μl of plasmid DNA, and place on ice for 30min; ③ Heat-shock introduction: Place at 42°C for 90s; ④ Membrane pore contraction: Ice bath for 2min; ⑤ Recovery culture: Add 800μl of liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1h (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 30min (the agar surface must be thoroughly dried), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium (LB with the corresponding antibiotic), culture with shaking for 12-16h, and extract the plasmid according to experimental needs.
Reference Application	pLV3-CMV-MCS-EF1a-mTagBFP2-Puro is a lentiviral expression vector that uses the strong CMV promoter to drive efficient expression of the target gene in the multiple cloning site (MCS); Meanwhile, the independent EF1a promoter drives mTagBFP2 blue fluorescent protein as an independent tracking marker, and it carries a Puromycin resistance gene for screening stably transfected cell lines.
Sharing Method	Public welfare sharing

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