title: Contamination of diverse nifH and nifH-like DNA into commercial PCR primers.
abstract: False-positive results due to DNA contamination in PCR reagents have become a big problem in the amplification of small amounts of DNA. Recently, it was revealed that PCR reagents were contaminated with the nifH (dinitrogenase reductase) gene. We found that the PCR primers supplied by some manufacturers contained nifH gene and nifH-like DNA. This contamination resulted in false-positive results when searching for nifH genes in environmental samples. The sequences of the contaminating DNA appeared to be widely varied in the phylogenetic analysis of nifH. For this reason, great care should be taken when analyzing trace amounts of nucleotides.
authors.affiliation: Department of Applied Biological Chemistry, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Goto
authors.forename: Shotaro
authors.initials: S
authors.lastname: Ando
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Hachisuka
authors.forename: Tadakatsu
authors.initials: T
authors.lastname: Yoneyama
mesh_terms: D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D017931:DNA Primers / Q000737:chemistry*; D005189:False Positive Reactions; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D004247:DNA; D010088:Oxidoreductases; C029130:nitrogenase reductase