Efficient removal of PCR inhibitors using agarose-embedded DNA preparations.
The use of agarose blocks containing embedded DNA improves the PCR amplification from templates naturally contaminated with polysaccharides or humic acids, two powerful PCR inhibitors. Presumably, the difference in size between the DNA macromolecules and these contaminants allows their effective removal from the agarose blocks by diffusion during the washing steps, whereas genomic DNA remains trapped within them. In addition, agarose-embedded DNA can be directly used for PCR since low melting point agarose does not interfere with the reaction. This simple and inexpensive method is also convenient for genomic DNAs extracted by other procedures, and it is potentially useful for samples containing other kinds of soluble inhibitors, overcoming this important problem of current amplification techniques.
Moreira
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Laboratoire de Biologie Cellulaire BC4, URA CNRS 2227, Université Paris-Sud, Bâtiment 444, 91405 Orsay Cedex, France. david.moreira@bc4.u-psud.fr
D004247:DNA / Q000737:chemistry*; D006812:Humic Substances / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D011134:Polysaccharides / Q000302:isolation & purification*; D012685:Sepharose / Q000737:chemistry*
D006812:Humic Substances; D011134:Polysaccharides; D004247:DNA; D012685:Sepharose