Jg. Hacia et al., 2-COLOR HYBRIDIZATION ANALYSIS USING HIGH-DENSITY OLIGONUCLEOTIDE ARRAYS AND ENERGY-TRANSFER DYES, Nucleic acids research, 26(16), 1998, pp. 3865-3866
High density oligonucleotide arrays (DNA chips) have been used in two
color mutational analysis of the 3.43 kb exon 11 of the hereditary bre
ast and ovarian cancer gene BRCA1. Two color analysis allows competiti
ve hybridization between a reference standard and an unknown sample, i
mproving the performance of the assay. Fluorescein and phycoerythrin d
yes were previously used due to their compatibility with a single line
488 nm excitation source. Here we show that an alternative dye combin
ation, containing the energy transfer dye system phycoerythrin.cy5 alo
ng with phycoerythrin, provides more evenly matched signal intensities
and decreased spectral overlap between the two fluorophores, while ma
intaining compatibility with a 488 nm excitation source.