Two color hybridization analysis using high density oligonucleotide arrays and energy transfer dyes.
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.
National Human Genome Research Institute, Building 49/3A14, National Institutes of Health, Bethesda, MD 20892, USA.
J G
JG
Hacia
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K
Edgemon
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B
Sun
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Stern
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Fodor
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Collins
D001483:Base Sequence; D004396:Coloring Agents; D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons; D005260:Female; D005456:Fluorescent Dyes; D019398:Genes, BRCA1; D006579:Heterozygote; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000235:genetics*; D010799:Phycoerythrin; D011110:Polymorphism, Genetic
D004396:Coloring Agents; D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D010799:Phycoerythrin