Rapid 16S ribosomal DNA sequencing from a single colony without DNA extraction or purification

Biotechniques. 1991 Jul;11(1):40-4.

Abstract

Ribosomal RNA sequences are useful for establishing phylogenetic relationships, for oligonucleotide probes and for characterization of uncultured organisms. We describe rapid ribosomal DNA sequencing using PCR with transcript sequencing. Nucleic acid specificity at three steps (amplification, transcription and sequencing) eliminated the need for nucleic acid extraction or purification. Sequence was obtained from a crude lysate from a single colony of bacteria. The basic sequencing method should be adaptable to provide rapid sequence information in a wide variety of applications.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Autoradiography
  • Base Sequence*
  • DNA, Bacterial / isolation & purification
  • DNA, Ribosomal*
  • Deoxyribonucleotides
  • Escherichia coli / genetics
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • RNA, Ribosomal, 16S / genetics*
  • RNA-Directed DNA Polymerase
  • Transcription, Genetic

Substances

  • DNA, Bacterial
  • DNA, Ribosomal
  • Deoxyribonucleotides
  • RNA, Ribosomal, 16S
  • RNA-Directed DNA Polymerase