Two color hybridization analysis using high density oligonucleotide arrays and energy transfer dyes

Nucleic Acids Res. 1998 Aug 15;26(16):3865-6. doi: 10.1093/nar/26.16.3865.

Abstract

High density oligonucleotide arrays (DNA chips) have been used in two color mutational analysis of the 3.43 kb exon 11 of the hereditary breast and ovarian cancer gene BRCA1 . Two color analysis allows competitive hybridization between a reference standard and an unknown sample, improving the performance of the assay. Fluorescein and phycoerythrin dyes werepreviously used due to their compatibility with a single line 488 nm excitation source. Here we show that an alternative dye combination, containing the energy transfer dye system phycoerythrin*cy5 along with phycoerythrin, provides more evenly matched signal intensities and decreased spectral overlap between the two fluorophores, while maintaining compatibility with a 488 nm excitation source.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Coloring Agents*
  • DNA Mutational Analysis / methods*
  • Exons
  • Female
  • Fluorescent Dyes
  • Genes, BRCA1*
  • Heterozygote
  • Humans
  • Nucleic Acid Hybridization / methods*
  • Oligodeoxyribonucleotides / genetics*
  • Phycoerythrin
  • Polymorphism, Genetic

Substances

  • Coloring Agents
  • Fluorescent Dyes
  • Oligodeoxyribonucleotides
  • Phycoerythrin