pLV2-CMV-MCS-IRES-TurboRFP-Puro - 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 sterile water to the plasmid dry powder to dissolve, 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°C 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 on a shaker at 37°C for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of recovered cells onto an LB plate with the corresponding antibiotic, keep the plate 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 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	pLV2-CMV-MCS-IRES-TurboRFP-Puro is a lentiviral expression vector that uses the strong CMV promoter to drive expression of the target gene in the multiple cloning site (MCS), and connects the TurboRFP (red fluorescent protein) and Puromycin resistance gene through an IRES (internal ribosome entry site) element, achieving co-expression of the target gene, red fluorescent tracing, and resistance selection.
Sharing Method	Public welfare sharing

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