pUC19-35S-FLAG-RBS - 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, and let stand at room temperature for 1min; ② Mixing: Mix 200μL competent cells 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 at 37℃ on a shaker for 1h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100μL) of recovered cells onto LB plates with the corresponding antibiotic resistance. Leave the plates upright for 30min (make sure the agar surface is completely dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick single colonies into 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	pUC19-35S-FLAG-RBS is a plasmid used for plant protein expression, carrying the 35S promoter, FLAG tag, and RBS. It has ampicillin resistance, with pUC19 as the backbone vector, and undergoes high-copy amplification in DH5α.
Sharing Method	Public welfare sharing