Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4045

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Purification of RNA from Cells and Tissues by Acid Phenol-Guanidinium Thiocyanate-Chloroform Extraction

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 100 words of the full text of this article appear below.


INTRODUCTION

In this single-step technique, cells are homogenized in guanidnium thiocyanate and the RNA is purified from the lysate by extraction with phenol:chloroform at reduced pH. Many samples can be processed simultaneously and speedily. The yield of total RNA depends on the tissue or cell source and is generally in the range of 4-7 µg/ml starting tissue or 5-10 µg/106 cells. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O.


MATERIALS

caution Chloroform:isoamyl alcohol (49:1, v/v)

Ethanol

Optional, please see Step 5.

recipe caution Formamide (Optional)

Deionized formamide is used for the storage of RNA.

Isopropanol

caution Liquid nitrogen

Mammalian cells

. . . [Full Text of this Article]


METHOD


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