pLX304-FLAG-MCS-NES - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24h; Aerobic
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20μL of sterile water to the plasmid lyophilized powder to dissolve it, and let it stand at room temperature for 1min; ② Mixing: Mix 200μl of competent cells with 5-10μl of plasmid DNA evenly, and place on ice for 30min; ③ Heat-shock introduction: Let stand at 42°C for 90s; ④ Membrane pore contraction: Ice bath for 2min; ⑤ Recovery culture: Add 800μl of liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1h (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 30min (make sure the agar surface is thoroughly dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid as needed for the experiment.
Reference Application	pLX304-FLAG-MCS-NES is a lentiviral expression vector that uses the CMV promoter to drive expression of the target gene in the multiple cloning site (MCS), with an N-terminal FLAG tag fused for easy detection, and a nuclear export signal (NES) linked to the C-terminus of the protein, localizing the fusion protein in the cytoplasm; The vector carries a Blasticidin resistance gene for selection of stably transfected cell lines.
Sharing Method	Public welfare sharing

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