pLV3-CMV-YPet-3×HA-EF1a-Blast - 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 sterile water to the plasmid dry powder to dissolve, 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 according to experimental needs.
Intended Use	pLV3-CMV-YPet-3×HA-EF1a-Blast is a lentiviral expression vector that uses the CMV promoter to drive expression of the YPet yellow fluorescent protein fused with a 3×HA tag, while the EF1a promoter independently drives the Blasticidin resistance gene for selection of stable cell lines.
Sharing Method	Public welfare sharing