pHIV-Luciferase-P2A-GFP - 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 of sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1 min; ② Mixing: Take 200 μL of competent cells and mix with 5-10 μL of 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 of 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 the plates upright for 30 min (ensure the agar surface is thoroughly dry before use), 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.
Reference Application	pHIV-Luciferase-P2A-GFP is a lentiviral (HIV-1 backbone) reporter gene vector that uses the CMV or EF1a promoter to drive co-expression of firefly luciferase (Luciferase) and green fluorescent protein (GFP) via the P2A self-cleaving peptide, enabling dual monitoring through bioluminescence (Luciferase quantification) and fluorescence (GFP tracing); The vector carries a Puromycin or Neomycin resistance gene for selection of stably transfected cell lines.
Sharing Method	Public welfare sharing

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