pLV3-4XIRF5-miniP-tdTomato-4×ISRE-miniP-mTagBFP2-Blast-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 of sterile water to the lyophilized plasmid powder and let stand at room temperature for 1 min; ② Mixing: Mix 200 μL of competent cells with 5-10 μL of plasmid DNA and place on ice for 30 min; ③ Heat-shock transformation: Place at 42℃ for 90 s; ④ Membrane pore contraction: Place on ice for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube and incubate at 37℃ with shaking at 150 r/min for 1 h; 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 thoroughly dried first), then invert and incubate for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic and culture with shaking for 12-16 h. Extract the plasmid as required for the experiment.
Reference Application	pLV3-4×IRF5-miniP-tdTomato-4×ISRE-miniP-mTagBFP2-Blast is a dual luciferase reporter lentiviral vector used for simultaneous monitoring of two independent interferon signaling pathways.
Sharing Method	Public Welfare Sharing