pLV3-CMV-EGFP-3×Myc-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 hours； Aerobic；
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20 μL of sterile water to the plasmid dry powder to dissolve, and let 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 introduction: Let stand at 42°C for 90 s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of recovered cells onto an LB plate with the corresponding antibiotic, keep the plate upright for 30 min (the agar surface must be completely dry first), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick a single monoclonal colony into liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid as needed for the experiment.
Reference Application	pLV3-CMV-EGFP-3×Myc-Puro is a lentiviral expression vector that uses the CMV promoter to drive the fusion expression of the EGFP green fluorescent protein and the 3×Myc tag, and carries a puromycin resistance gene for screening stable cell lines.
Sharing Method	Public welfare sharing