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Purification of RNA and DNA Introducing DNA into cells

Purification of RNA and DNA Introducing DNA into cells. May 2. Purification of DNA. Plasmid DNA (1-6,9,15) alkaline lysis commercial kits DNA from mammalian cells (1-18) cultured cells, tissues, blood DNA from yeast cells (12-8). Concentrating DNA. Ethanol precipitation.

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Purification of RNA and DNA Introducing DNA into cells

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  1. Purification of RNA and DNAIntroducing DNA into cells May 2

  2. Purification of DNA • Plasmid DNA (1-6,9,15) • alkaline lysis • commercial kits • DNA from mammalian cells (1-18) • cultured cells, tissues, blood • DNA from yeast cells (12-8)

  3. Concentrating DNA • Ethanol precipitation Add 2X volume of 95-100% ethanol

  4. Purification of RNA • RNA composition of a cell • Ribonucleases (4-1)

  5. Introduction of DNA into cells • bacterial cells • competent cells (E.coli) (1-1) • electroporation (1-4) • cultured mammalian cells • calcium phosphate (9-5) • electroporation • commercial lipid reagents • viral vectors

  6. Today • immunoblot of transferred proteins • transfection of vero cells (to prepare positive and negative controls for quantitative real-time PCR)

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