Successful pyrosequencing of GC-rich DNA sequences by partial substitution of deoxyguanosine with deoxyinosine

Diagn Mol Pathol. 2010 Jun;19(2):123-5. doi: 10.1097/PDM.0b013e3181b5a2d5.

Abstract

We experienced significant problems while developing a PyrosequencingTM (Biotage, Uppsala, Sweden) assay to characterize the rifampin resistance-determining region of Mycobacteriumtuberculosis, a target with high guanosine-cytosine content. This paper describes the successful use of a modified pyrosequencing protocol through partial substitution of deoxyguanosine triphosphate with deoxyinosine triphosphate in the polymerase chain reaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Base Composition
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism*
  • Deoxyguanosine / metabolism*
  • Humans
  • Inosine / analogs & derivatives*
  • Inosine / metabolism
  • Mycobacterium tuberculosis / genetics
  • Pathology, Molecular / methods*
  • Sequence Analysis, DNA / methods*
  • Specimen Handling / methods*
  • Sweden

Substances

  • ARR-2 protein, Bacteria
  • Bacterial Proteins
  • DNA, Bacterial
  • Inosine
  • Deoxyguanosine
  • deoxyinosine