Unit

UNIT 10.1 Purification and Concentration of DNA from Aqueous Solutions

  1. David Moore

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im1001s8

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Moore, D. 2001. Purification and Concentration of DNA from Aqueous Solutions. Current Protocols in Immunology. 8:I:10.1:10.1.1–10.1.9.

Author Information

  1. Massachusetts General Hospital, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: DEC 1993

Abstract

This unit presents basic procedures for manipulating solutions of single- or double-stranded DNA through purification and concentration steps. The Basic Protocol, using phenol extraction and ethanol precipitation, is appropriate for the purification of DNA from small volumes (<0.4 ml) at concentrations <1 mg/ml. Isopropanol may also be used to precipitate DNA, as described in an alternate protocol. Three support protocols outline methods to buffer the phenol used in extractions, concentrate DNA using butanol, and extract residual organic solvents with ether. An alternative to these methods is nucleic acid purification using glass beads, and is described here. These protocol may also be used for purifying RNA. The final protocols provide modifications to the Basic Protocol that are used for concentrating RNA and extracting and precipitating DNA from larger volumes and from dilute solutions, and for removing ow-molecular-weight oligonucleotides and triphosphates.