pT7-MCS-TEV-mCherry-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 for dissolution, and let stand at room temperature for 1 min; ② Mixing: Mix 200μL competent cells with 5-10μL plasmid DNA, and place on ice for 30 min; ③ Heat shock introduction: Let stand at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2 min; 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 (100μL) of recovered cells onto LB plates with the corresponding antibiotic, leave the plates upright for 30 min (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, culture with shaking for 12-16h, and extract the plasmid as needed for the experiment.
Reference Use	pT7-MCS-TEV-mCherry-10×His-Kan is a vector used for T7 prokaryotic expression in Escherichia coli, with a TEV protease cleavage site at the N-terminus, a C-terminal fusion of mCherry red fluorescent protein and a 10×His tag, and carries kanamycin resistance. It is suitable for visual expression and affinity purification of target proteins.
Sharing Method	Public welfare sharing