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Design and evaluation of PCR primers to amplify bacterial 16S ribosomal DNA fragments used for community fingerprinting

Tipo de material: TextoTextoSeries ; Journal of MicroBiological Methods, 44(3), p.253-262, 2001Trabajos contenidos:
  • Watanabe, K
  • Kodama, Y
  • Harayama, S
Tema(s): Recursos en línea: Resumen: Denaturing gradient gel electrophoresis of PCR-amplified 16S ribosomal DNAZrDNA.fragments has frequently been applied to the fingerprinting of natural bacterial populationsZPCRrDGGE.. In this study, sequences of bacterial universal primers frequently used in PCRrDGGE were compared with 16S rDNA sequences that represent recently proposed divisions in the domain Bacteria. We found mismatches in 16S rDNA sequences from some groups of bacteria. Inosine residues were then introduced into the bacterial universal primers to reduce amplification biases caused by these mismatches. Using the improved primers, phylotypes affiliated with Verrucomicrobia and candidate division OP11, were detected in DGGE fingerprints of groundwater populations, which have not been detected by PCRrDGGE with conventional universal primers.
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Documentos solicitados Documentos solicitados CICY Documento préstamo interbibliotecario Ref1 B-11191 (Browse shelf(Opens below)) Available

Denaturing gradient gel electrophoresis of PCR-amplified 16S ribosomal DNAZrDNA.fragments has frequently been applied to the fingerprinting of natural bacterial populationsZPCRrDGGE.. In this study, sequences of bacterial universal primers frequently used in PCRrDGGE were compared with 16S rDNA sequences that represent recently proposed divisions in the domain Bacteria. We found mismatches in 16S rDNA sequences from some groups of bacteria. Inosine residues were then introduced into the bacterial universal primers to reduce amplification biases caused by these mismatches. Using the improved primers, phylotypes affiliated with Verrucomicrobia and candidate division OP11, were detected in DGGE fingerprints of groundwater populations, which have not been detected by PCRrDGGE with conventional universal primers.

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