pLV3-CMV-EBFP-MCS-Puro - 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 to dissolve it, 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 on a shaker for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of the recovered cells onto LB plates with the corresponding antibiotic. Leave the plates upright for 30 min (the agar surface must be dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid as required for the experiment.
Reference Use	pLV3-CMV-EBFP-MCS-Puro is a lentiviral expression vector that uses the strong CMV promoter to drive fusion expression of EBFP (enhanced blue fluorescent protein) with the target gene in the multiple cloning site (MCS), facilitating fluorescence tracing and localization of the target protein; It also carries a Puromycin resistance gene for screening stably transfected cell lines.
Sharing Method	Public welfare sharing

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