pLV3-CMV-MCS-EF1a-mScarlet3-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 sterile water to the plasmid dry powder for dissolution, and let it stand at room temperature for 1min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA and place on ice for 30min; ③ Heat-shock introduction: Let stand at 42°C 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°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 (ensure the agar surface is thoroughly dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid according to experimental needs.
Reference Application	pLV3-CMV-MCS-EF1a-mScarlet3-Puro is a lentiviral expression vector that uses the strong CMV promoter to drive expression of the target gene cloned into the multiple cloning site (MCS), while the EF1a promoter independently drives the mScarlet3 red fluorescent protein as a tracing marker. It also carries a Puromycin resistance gene for selection of stable cell lines.
Sharing Method	Public welfare sharing