pLV3-EF1a-Tdtomato-Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	37℃; 18-24h; Aerobic
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20μL sterile water to the plasmid powder for dissolution, and let stand at room temperature for 1min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, and place on ice for 30min; ③ Heat shock introduction: Let stand at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture at 37℃ on a shaker for 1h (150 r/min); ⑥ Screening culture: Spread an appropriate volume (100μl) of recovered cells on an LB plate with the corresponding resistance, leave the plate upright for 30min (the agar surface must be dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick a single colony into LB liquid Recommended Medium with the corresponding resistance, culture with shaking for 12-16h, and extract the plasmid as needed for the experiment.
Reference Use	pLV3-EF1a-tdTomato is a lentiviral vector that uses the EF1a promoter to drive stable expression of the tdTomato red fluorescent protein in mammalian cells, for fluorescence tracing or labeling of transduced cells, and often carries Puromycin resistance for screening stable cell lines.
Sharing Method	Public Welfare Sharing

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