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Molecular Pathology

DNA purification

PCR Inhibitors


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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● PCR inhibitors are commonly encountered; removing these containments is vital to insure good PCR amplification
● The chemical structure of PCR inhibitors and mechanism(s) of inhibition show enormous variation
● Inhibitors include excess salts such as KCl and NaCl, detergents/hydrophobic chemicals (phenol, SDS, chloroform, sodium deocyholate, and sarkosyl), inappropriate heating, loss of reagents required for PCR (such as Mg++, Taq polymerase denaturation, etc), alcohols (ethanol and isopropyl alcohol), heparin, bromophenol blue, agarose, inosine, humic acids, deoxyuridine, excess nucleic acids in the amplification reaction, various polysaccharides (OpenWetWare)
● Since inhibitors can be either hydrophilic or hydrophobic (or both), DNA purification purification protocols usually involve subjecting the crude nucleic acid mix to both hydrophilic and hydrophobic washing steps

End of Molecular Pathology > DNA Purification > PCR Inhibitors


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