(1) Adéle Mennerat, associate professor at the University of Bergen, and Ben C. Sheldon, professor at Oxford University, in their microbial ecology article on PCR contamination “How to Deal with PCR Contamination in Molecular Microbial Ecology” (2014) suggests that a new standard method of purifying PCR reagents with the endonuclease restriction enzyme Sau3AI be used. (2) The authors show through past literature as well as experiments that reagents contribute contaminants even when extreme care is taken, so the reagents must be purified before the template DNA is added to avoid misrepresentations of community structure. (3) The author’s purpose is to prove with OTU results that the best way to purify PCR reagents is with Sau3AI followed by a heat treatment to irreversibly inactivate the enzyme in order to protect the template DNA from enzyme degradation, which will mean the resulting product is of only the community DNA. (4) The intended audience is ecologists who study microbial community structure, as well as all researchers who use PCR, and the relationship the authors create is friendly and cooperative in hopes that other researchers will agree with the value of enzyme purification.
Citation: Mennerat A, Sheldon BC. 2014. How to Deal with PCR Contamination in Molecular Microbial Ecology. Microb Ecol. 68:834-841. DOI: 10.1007/s00248-014-0453-7y
