Identification of mixed bacterial DNA contamination in broad-range PCR amplification of 16S rDNA V1 and V3 variable regions by pyrosequencing of cloned amplicons.

@article{Grahn2003IdentificationOM,
  title={Identification of mixed bacterial DNA contamination in broad-range PCR amplification of 16S rDNA V1 and V3 variable regions by pyrosequencing of cloned amplicons.},
  author={Niclas Grahn and Margaretha Olofsson and Katarina Ellnebo-Svedlund and Hans J{\"u}rg Monstein and Jon Jonasson},
  journal={FEMS microbiology letters},
  year={2003},
  volume={219 1},
  pages={87-91}
}
Using a sensitive and rapid method combining broad-range PCR amplification of bacterial 16S rDNA fragments and pyrosequencing for detection, identification and typing, we have found contaminating bacterial DNA in our reagents used for PCR. Identified bacteria are the water-borne bacterial genera Pseudomonas, Stenotrophomonas, Xanthomonas, Ralstonia and Bacillus. Our results are in concordance with recent reports of contaminated industrial water systems. In light of this conclusion, we believe… CONTINUE READING