pT7-MCS-TEV-EGFP-10×His-Kan-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 to dissolve, let stand at room temperature for 1min; ② Mixing: Take 200μl competent cells and 5-10μl plasmid DNA, mix well, 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, 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, keep plates upright for 30min (the agar surface must be dry first), 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 plasmid according to experimental needs.
Reference Application	pT7-MCS-TEV-EGFP-10×His-Kan is a T7 promoter prokaryotic expression vector used in Escherichia coli, designed for fusion expression, visual tracking, and affinity purification of target proteins.
Sharing Method	Public welfare sharing