pGreen II-62-SK-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: Mix 200μl competent cells with 5-10μl plasmid DNA, 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 at 37℃ on a shaker for 1h (150 r/min); ⑥ Screening culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding antibiotic, leave plates upright for 30min (the agar surface must be 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 according to experimental needs.
Reference Use	pGreen II-62-SK is a plant binary expression vector used for plant cells, carrying the CaMV 35S promoter, multiple cloning site (MCS), and kanamycin resistance, suitable for overexpression and functional studies of plant genes.
Sharing Method	Public welfare sharing