pLV2-MPC-24×GCN4-Puro - 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 dry powder for dissolution, and let stand at room temperature for 1min； ② Mixing: Take 200μl competent cells and mix 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 on a shaker at 37℃ for 1h (150 r/min)； ⑥ Selection culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding antibiotic resistance. Leave the plates upright for 30min (the agar surface must be completely dry), then invert and culture for 12-16h until colonies appear； ⑦ Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic resistance, culture with shaking for 12-16h, and extract the plasmid as required for the experiment.
Reference Application	pLV2-MPC-24×GCN4-Puro is a lentiviral expression vector that drives the expression of 24 tandem repeats of the GCN4 peptide (a transcriptional activation domain derived from yeast) fused to a target protein via the CMV promoter. It is used for transcriptional activation or protein recruitment experiments, and carries a Puromycin resistance gene for selection of stable cell lines.
Sharing Method	Public welfare sharing