Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4455

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Purification of Nucleic Acids by Extraction with Phenol:Chloroform

Joseph Sambrook and David W. Russell

This protocol was adapted from "Commonly Used Techniques in Molecular Cloning," Appendix 8, in Molecular Cloning, Volume 3, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001.

The first 15% of the full text of this article appears below.


INTRODUCTION

This protocol describes the standard method for nucleic acid purification by extraction first with phenol:chloroform (optionally containing hydroxyquiniline at 0.1%) and then with chloroform to remove any remaining phenol. The procedure takes advantage of the fact that deproteinization is more efficient when two different organic solvents are used instead of one.


MATERIALS

Reagents

caution Chloroform

Ethanol

caution Ether (optional)

Nucleic acid solution to be purified

caution caution Phenol:Chloroform (1:1)

recipe TE (pH 7.8) . . . [Full Text of this Article]

Equipment


METHOD

TROUBLESHOOTING


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Standard Ethanol Precipitation of DNA in Microcentrifuge Tubes
Joseph Sambrook and David W. Russell
CSH Protocols 2006: 4456. [Extract] [Full Text]



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