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Molecular Pathology

DNA Purification

Silica Adsorption


Author: Rodney E. Shackelford, D.O., Ph.D. (see Reviewers page)
Revised: 14 April 2012, last major update April 2012
Copyright: (c) 2008-2011, PathologyOutlines.com, Inc.

Definition
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● Uses a silica micro-channel matrix with a specific salt concentration and pH (Wikipedia)
● DNA within a previously prepared cell lysate or other crude nucleic acid mixture preferentially binds to the walls of the micro-channels in the presence of a high salt buffer
● The DNA binds to the silica under conditions of high ionic strength and a pH at or below the pKa of the silica surface silanol (SiOH) groups
● Multiple washing steps with an alcohol-based wash buffer remove other cellular constituents, particularly proteins
● Later the pure DNA is removed with a specific low-salt elution buffer; presently, this technique is relatively expensive, limiting its application
● Many automated DNA extraction machines employ a silica matrix for DNA capture and purification

End of Molecular Pathology > DNA Purification > Silica Adsorption


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