Development of a modified method for sequencing high G:C regions in chicken anemia virus genome

J Biochem Mol Biol Biophys. 2002 Jun;6(3):229-32. doi: 10.1080/10258140290022310.

Abstract

Two areas in the chicken anemia virus (CAV) genome have high G:C content with secondary structures. These two G:C rich areas could not be sequenced with Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit. Several modifications were carried out to solve the problem. Finally, a package of modified method was developed to sequence the high G:C areas. The result showed that the Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit with the normal procedures are not suitable for sequencing the high G:C regions of the CAV genome. The present developed method made the Perkin Elmer's Kit useful for the first time to sequence the G:C rich hairpin structures of the CAV genome. The system may be useful to sequence all other G:C rich DNA templates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Chicken anemia virus / genetics*
  • Cloning, Molecular
  • Cytosine
  • Genetic Techniques*
  • Genome, Viral*
  • Guanine
  • Models, Genetic
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA / methods*
  • Tumor Cells, Cultured

Substances

  • Guanine
  • Cytosine