pLV3-CMV-Gln-3×FLAG-8×His-Puro - Basic Information

Recommended Medium	Yeast extract: 5.0 g, Peptone: 10.0 g, NaCl: 10.0 g, pH: 7.0 (25℃)
Growth Conditions	37℃； 18-24 h； Aerobic
Storage Conditions	2-8℃
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℃ 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 on a shaker at 37℃ for 1 h (150 r/min)； ⑥ Selection culture: Spread an appropriate volume (100 μL) of recovered cells onto LB plates with the corresponding resistance. Leave the plates upright for 30 min (the agar surface must be thoroughly dry), then invert and culture for 12-16 h until colonies appear； ⑦ Extraction: Pick single monoclonal colonies into liquid LB Recommended Medium with the corresponding resistance, culture with shaking for 12-16 h, and extract the plasmid according to experimental needs.
Reference Application	pLV3-CMV-Gln-3×FLAG-8×His-Puro is a lentiviral expression vector that uses the strong CMV promoter to drive glutaminase (Gln) gene expression, fusing a 3×FLAG tag and an 8×His tag to the N-terminus or C-terminus of the target protein (for efficient detection and purification), and employs the Puromycin resistance gene to select stably transfected cell lines.
Sharing Method	Public welfare sharing