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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© 2005 Humana Press Inc.
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Ian, B.M. (2005). Extraction of RNA From Cells and Tissue. In: Fennell, J.P., Baker, A.H. (eds) Hypertension. Methods In Molecular Medicine™, vol 108. Humana Press. https://doi.org/10.1385/1-59259-850-1:139
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DOI: https://doi.org/10.1385/1-59259-850-1:139
Publisher Name: Humana Press
Print ISBN: 978-1-58829-323-7
Online ISBN: 978-1-59259-850-2
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