pLV2-EGFP-mRFP1-Puro-Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	37℃; 18-24 hours; Aerobic;
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder for dissolution, and let stand at room temperature for 1 min; ② Mixing: Take 200μl competent cells and mix with 5-10μl plasmid DNA, and place on ice for 30 min; ③ Heat shock introduction: Let stand at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture on a shaker at 37℃ for 1 h (150 r/min); ⑥ Screening culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding resistance, place 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 resistance, culture with shaking for 12-16 h, and extract the plasmid according to experimental needs.
Reference Use	Used for simultaneous expression of both EGFP and mRFP1 fluorescent proteins in mammalian cells, and it is a lentiviral vector carrying a puromycin resistance gene.
Sharing Method	Public welfare sharing