title: Extraction of RNA from cells and tissue.
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.
authors: Department of Obstetrics and Gynecology, University of Wisconsin, Madison, USA.
authors: Ian M
authors: IM
authors: Bird
mesh_terms: D002478:Cells, Cultured; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility; D013053:Spectrophotometry
chemical_list: D012313:RNA