HiYield Plasmid Mini Kit 3.0<\/strong>\u00a0is specially designed for rapid isolation of plasmid or cosmid DNA from 1-4 ml of bacterial cultured cells. As high as 50 \u00b5g of high-quality plasmid DNA can be purified in less than 30 minutes and is ready for restriction digestion, ligation, PCR, and sequencing reaction. No phenol extraction or alcohol precipitation is required in this protocol. In the process, clear and extra pure cell lysate with minimal genomic DNA and RNA contaminants can be obtained through the modified alkaline lysis method and RNase treatment. In the presence of a chaotropic salt, the plasmid DNA within the lysate will then bind to the glass fiber matrix equipped in the spin column. The contaminants are washed away with an ethanol-containing wash buffer. Finally, the purified plasmid DNA is eluted by a low salt elution buffer or distilled water. Typical yields of high-purity are 20~50 \u00b5g for high-copy number plasmids or 3~10 \u00b5g for low-copy-number plasmids. <\/p>\n<\/div>\n<\/div>\n Restriction enzyme digestion, library screening, ligation, PCR,\u00a0transformation\/sequencing reactions<\/p>\n<\/div>\n<\/div>\n The quality of\u00a0HiYield Plasmid Mini Kit 3.0\u00a0<\/strong>is tested on a lot-to-lot basis. The kit is tested by isolation of plasmid DNA from 5 ml culture of\u00a0E.coli<\/em>\u00a0DH5\u03b1 transformed with the plasmid pGAD424 (A600<\/sub>> 2 units\/ml). More than 40 \u00b5g of plasmid DNA should be obtained. One \u00b5g of the purified product is also tested for restriction enzyme digested with\u00a0EcoR<\/em>\u00a0I followed by agarose gel analysis.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n Store at<\/span><\/p>\n # FYG011-100P<\/b>\u00a0(100 preps)<\/span><\/p>\n
HiYield Plasmid Mini Kit 3.0 improves the structure of the membrane and the composed of glass fiber makes\u00a0the purified plasmids purer and the lower concentration of endotoxin<\/strong>.\u00a0<\/p>\n\n
1 – 4 mL of bacterial cells<\/li>\n<\/ul>\n\n
spin column (centrifuge)<\/li>\n<\/ul>\n\n
up to 50 \u03bcg of plasmid\/cosmid DNA<\/li>\n<\/ul>\n\n
up to 30 minutes<\/li>\n<\/ul>\n\n
50 – 100 \u03bcL<\/li>\n<\/ul>\n<\/div>\n<\/div>\n![\"\"](\"https:\/\/si.secda.info\/yb_md\/wp-content\/uploads\/2017\/07\/RT-300x222.png\")
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