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      Extraction of RNA from cells and tissue.

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      Methods in molecular medicine

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          Abstract

          Total undegraded cellular RNA, largely free of contaminating DNA, can be rapidly and easily isolated from homogenized tissue or cultured cells using acid pH guanidinium thiocyanate/phenol/chloroform extraction. Commercially available acid phenol/guanidinium thiocyanate solutions allow one-step complete solubilization of tissue or cells under conditions that inhibit RNAse activity. We describe here a detailed procedure for total RNA extraction from homogenized tissues or cultured cells using such reagents, together with additional purification steps by passage through Phase-Lock gel tubes, and recommend optional chromatographic poly(A+) RNA enrichment procedure using Mini Oligo(dT) spin columns, together with the spectrophotometric determination of RNA yield and purity.

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          Author and article information

          Journal
          Methods Mol. Med.
          Methods in molecular medicine
          1543-1894
          1543-1894
          2005
          : 108
          Affiliations
          [1 ] Department of Obstetrics and Gynecology, University of Wisconsin, Madison, USA.
          16028681

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