Contamination of diverse nifH and nifH-like DNA into commercial PCR primers.
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.
Goto
Masahiro
M
Department of Applied Biological Chemistry, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Ando
Shotaro
S
Hachisuka
Yusuke
Y
Yoneyama
Tadakatsu
T
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
D017931:DNA Primers; D004247:DNA; D010088:Oxidoreductases; C029130:nitrogenase reductase