Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

Table of Contents

Expand All | Contract All

A Single-Step Method for the Simultaneous Preparation of DNA, RNA, and Protein from Cells and Tissues

(Protocol summary only for purposes of this preview site)

The following protocol (Chomczynski 1993) allows the simultaneous recovery of RNA, DNA, and protein from an aliquot of tissue or cells. Like its predecessor (Chomczynski and Sacchi 1987), this method involves lysis of cells with a monophasic solution of guanidine isothiocyanate and phenol. Addition of chloroform generates a second (organic) phase into which DNA and proteins are extracted, leaving RNA in the aqueous supernatant. The DNA and proteins can be isolated from the organic phase by sequential precipitation with ethanol and isopropanol, respectively. The DNA recovered from the organic phase is 20 kb in size and is a suitable template for PCRs. The proteins, however, remain denatured as a consequence of their exposure to guanidine and are used chiefly for immunoblotting. The RNA precipitated from the aqueous phase with isopropanol can be further purified by chromatography on oligo(dT)-cellulose columns and/or used for northern blot hybridization, reverse transcription, or RT-PCRs.


Previous   |   Next

Save 30% & Get Free Shipping!

Save 30% at checkout on our website.
(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

Search for information about other protocols included in the book:

Read What Others Are Saying About Molecular Cloning:

Tom Maniatis wins Lasker

'Not Quite a Book Prize'

Get alerts about special offers from Cold Spring Harbor Laboratory Press free to your Inbox