pLV-EF1a-TagRFP-EGFP-IRES-BSD - 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 it, and let it stand at room temperature for 1 min; ② Mixing: Take 200 μL of competent cells and mix with 5-10 μL of plasmid DNA, then place on ice for 30 min; ③ Heat shock introduction: Let stand at 42°C for 90 s; ④ Membrane pore contraction: Place on ice bath for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of the recovered cells onto LB plates with the corresponding antibiotic resistance, leave the plates upright for 30 min (the agar surface must be absolutely dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic resistance, culture with shaking for 12-16 h, and extract the plasmid as required by the experiment.
Reference Use	pLV-EF1a-TagRFP-EGFP-IRES-BSD is a dual-fluorescence reporter lentiviral vector for mammalian cells. It uses the EF1a promoter to drive expression of TagRFP and EGFP fusion protein, and is coupled with the blasticidin resistance gene through an IRES element for stable cell line selection.
Sharing Method	Public welfare sharing