pLV-attB4-CMV-MCS-EF1a-Puro-Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25°C)
Growth Conditions	37°C； 18-24 hours； Aerobic；
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1 min； ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA and place on ice for 30 min； ③ Heat shock introduction: Place at 42°C for 90 s； ④ Membrane pore contraction: Ice bath for 2 min； ⑤ Recovery culture: Add 800μl 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 on LB plates with corresponding resistance, leave the plates upright for 30 min (ensure the agar surface is completely dry), then invert and culture for 12-16 h until colonies appear； ⑦ Extraction: Pick single colonies into liquid Recommended Medium with corresponding resistance, culture with shaking for 12-16 h, and extract plasmid according to experimental needs
Reference Application	pLV-attB4-CMV-MCS-EF1a-Puro is a lentiviral vector that uses the CMV promoter to drive expression of the target gene in the multiple cloning site (MCS), and uses the EF1a promoter to drive the puromycin resistance gene for selection of stable cell lines.
Sharing Method	Public Welfare Sharing