Abstract
To improve signal detection on cDNA microarrays, we adapted a fluorescent oligonucleotide dendrimeric signal amplification system to microarray technology. This signal detection method requires 16-fold less RNA for probe synthesis, does not depend on the incorporation of fluorescent dNTPs into a reverse transcription reaction, generates a high signal-to-background ratio, and can be used to allow for multichannel detection on a single chip. Furthermore, since the dendrimers can be detected individually, it may be possible, by employing dendrimer-binding standards, to calculate the numbers of bound cDNAs can be estimated. These features make the dendrimer signal detection reagent ideal for high-throughput functional genomics research.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Biotechnology / methods
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Carbocyanines / metabolism
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DNA Primers / chemistry*
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DNA Primers / genetics
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DNA Primers / metabolism
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DNA, Complementary / analysis
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DNA, Complementary / genetics
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Fluorescent Dyes / metabolism
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Gene Expression Profiling / methods
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Genomics / methods*
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Nucleic Acid Conformation*
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Nucleic Acid Hybridization / methods
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Oligodeoxyribonucleotides / chemistry*
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Oligodeoxyribonucleotides / genetics
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Oligodeoxyribonucleotides / metabolism
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Oligonucleotide Array Sequence Analysis / methods*
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RNA, Messenger / analysis
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RNA, Messenger / genetics
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Reference Standards
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Reverse Transcriptase Polymerase Chain Reaction
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Sensitivity and Specificity
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Substrate Specificity
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Transcription, Genetic / genetics
Substances
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Carbocyanines
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DNA Primers
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DNA, Complementary
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Fluorescent Dyes
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Oligodeoxyribonucleotides
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RNA, Messenger
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cyanine dye 3
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cyanine dye 5