pLV3-ARE-luc2P-EF1a-Neo - 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 h; Aerobic
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20 μL sterile water to the plasmid dry powder for dissolution 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 recovered cells onto LB plates with the corresponding antibiotic. Leave plates upright for 30 min (agar surface must be dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single colonies into liquid Recommended Medium containing the corresponding antibiotic and culture with shaking for 12-16 h. Extract plasmid as needed for the experiment.
Reference Application	pLV3-ARE-luc2P-EF1a-Neo is a lentiviral reporter vector that uses ARE (antioxidant response element) to drive expression of the luc2P (firefly luciferase) reporter gene for monitoring the activation level of the Nrf2/ARE signaling pathway (reflecting cellular oxidative stress status); At the same time, the EF1a promoter independently drives the Neo/G418 resistance gene for selection of stably transfected cell lines.
Sharing Method	Public Welfare Sharing