PACE analysis of the heteroduplexes formed between PCR-amplified 16S rRNA genes: Estimation of sequence similarity and rDNA complexity

Romilio T. Espejo, Carmen Gloria Feijóo, Jaime Romero, Mónica Vásquez

Research output: Contribution to journalArticlepeer-review

52 Citations (Scopus)

Abstract

Analysis of the 16S rRNA genes retrieved directly from different environments has proven to be a powerful tool that has greatly expanded our knowledge of microbial diversity and phylogeny. It is shown here that sequence similarity between 80 and 100% among 16S rDNAs can be estimated by the electrophoretic migration of their heteroduplexes. This was measured by hybridization and electrophoresis in polyacrylamide gels of the product obtained after PCR amplification of almost the entire 16S rRNA gene from different bacterial species. These heteroduplexes were also observed after amplification of samples containing DNA from two or more bacterial species and a procedure was applied to identify reliably heteroduplexes among the amplification products. The electrophoretic migration of the heteroduplexes observed after PCR was used to detect the presence of 16S rDNAs with different sequences in DNA extracted from both a mixture of two bacterial species and samples containing a natural bacterial community.

Original languageEnglish
Pages (from-to)1611-1617
Number of pages7
JournalMicrobiology
Volume144
Issue number6
DOIs
Publication statusPublished - Jun 1998

Keywords

  • 16s rRNA
  • Diversity
  • Heteroduplex
  • Phylogeny

ASJC Scopus subject areas

  • Microbiology

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