pLV2-TRE3GS-mCherry-EGFP-MCS-TetOne-Puro - 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 sterile water to the plasmid dry powder to dissolve it, and let stand at room temperature for 1 min; ② Mixing: Take 200 μL competent cells and mix with 5-10 μL plasmid DNA, and place on ice for 30 min; ③ Heat-shock introduction: Let stand at 42℃ for 90 s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800 μL liquid Recommended Medium to each tube, and culture at 37℃ with shaking 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 (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 LB Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid according to experimental needs.
Reference Application	pLV2-TRE3GS-mCherry-EGFP-MCS-TetOne-Puro is a Tet-On inducible lentiviral vector that uses the TRE3GS promoter to drive co-expression of mCherry, EGFP, and the gene of interest in the multiple cloning site (MCS) in the presence of doxycycline (Dox), with TetOne providing the transactivator, and it also carries puromycin resistance for selection of stable cell lines.
Sharing Method	Public welfare sharing

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