The pyrosequencing protocol for bacterial genomes

Methods Mol Biol. 2015:1231:49-75. doi: 10.1007/978-1-4939-1720-4_4.

Abstract

The pyrosequencing methodology was applied in 2005 by 454 Lifesciences to the emerging field of next generation sequencing (NGS), revolutionizing the way of DNA sequencing. In the last years the same strategy grew up and was technologically updated, reaching a high throughput in terms of amount of generated sequences (reads) per run and in terms of length of sequence up to values of 800-1,000 bases. These features of pyrosequencing perfectly fit to bacterial genome sequencing for the de novo assemblies and resequencing as well. The approaches of shotgun and paired ends sequencing allow the bacterial genome finishing providing a high-quality data in few days with unprecedented results.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apyrase / chemistry
  • Bacteria / genetics*
  • Chromosome Mapping
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics*
  • DNA-Directed DNA Polymerase / chemistry
  • Dideoxynucleotides / chemistry
  • Genome, Bacterial*
  • Genomic Library*
  • High-Throughput Nucleotide Sequencing / instrumentation
  • High-Throughput Nucleotide Sequencing / methods*
  • Luciferases / chemistry
  • Molecular Sequence Annotation
  • Polymerase Chain Reaction / methods
  • Sequence Analysis, DNA / statistics & numerical data
  • Sulfate Adenylyltransferase / chemistry

Substances

  • DNA, Bacterial
  • Dideoxynucleotides
  • Luciferases
  • Sulfate Adenylyltransferase
  • DNA-Directed DNA Polymerase
  • Apyrase