A new approach using multiplex long accurate PCR and yeast artificial chromosomes for bacterial chromosome mapping and sequencing

Genome Res. 1996 May;6(5):448-53. doi: 10.1101/gr.6.5.448.

Abstract

An efficient approach for structural studies on bacterial chromosomes is presented. It is based on high-resolution PCR map construction by using a multiplex long accurate PCR (MLA PCR) protocol and a YAC clone carrying the region to be mapped as indicator. The high-resolution PCR map of the bacillus subtilis rrnB-dnaB region is presented as an example. Data are also presented on the use of DNA generated by LA PCR for sequencing; they are relevant to LA PCR induced mutations and justify the application of such mapping for sequencing long stretches of bacterial chromosomes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / genetics
  • Bacterial Proteins / genetics
  • Base Sequence
  • Chromosome Mapping / methods*
  • Chromosomes, Artificial, Yeast / metabolism
  • Chromosomes, Bacterial*
  • DNA Helicases*
  • DNA, Bacterial
  • DnaB Helicases
  • Electrophoresis, Gel, Pulsed-Field
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods
  • Sequence Analysis, DNA / methods*
  • rRNA Operon

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA Helicases
  • DnaB Helicases

Associated data

  • GENBANK/D83243
  • GENBANK/D83244