Cross-Hybridization of Closely Related Genes on High-Density Macroarrays

DNA macroarrays are used in many areas of molecular biology research for applications ranging from gene discovery to gene expression profiling. As an increasing number of specialized macroarrays containing genes related by function or pathway are becoming available, a question that needs to be addre...

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Main Authors: N.A. Miller, Q. Gong, R. Bryan, M. Ruvolo, L.A. Turner, S.T. LaBrie
Format: Article
Language:English
Published: Taylor & Francis Group 2002-03-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/02323pf01
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author N.A. Miller
Q. Gong
R. Bryan
M. Ruvolo
L.A. Turner
S.T. LaBrie
author_facet N.A. Miller
Q. Gong
R. Bryan
M. Ruvolo
L.A. Turner
S.T. LaBrie
author_sort N.A. Miller
collection DOAJ
description DNA macroarrays are used in many areas of molecular biology research for applications ranging from gene discovery to gene expression profiling. As an increasing number of specialized macroarrays containing genes related by function or pathway are becoming available, a question that needs to be addressed is the level of hybridization signal specificity between highly similar genes that can be achieved. We have examined the ability of our LifeGrid™ macroarrays to distinguish hybridization signals between closely related genes. We determined the level of cross-hybridization among genes ranging from 52% to 94% sequence identity. Fragments of genes from five protein families were arrayed onto nylon filters. The filters were subsequently hybridized with a 33P-labeled probe prepared from a pool of synthetic mRNA transcripts containing a representative of each protein family. We found that fragments containing sequences with up to 94% sequence identity displayed relatively little cross-hybridization. We conclude that this macroarray system is very specific and that hybridization signals from closely related genes can be reliably measured.
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spelling doaj-art-a0343678dc7b432982fef0daa5093d9c2025-08-20T02:25:54ZengTaylor & Francis GroupBioTechniques0736-62051940-98182002-03-0132362062510.2144/02323pf01Cross-Hybridization of Closely Related Genes on High-Density MacroarraysN.A. Miller0Q. Gong1R. Bryan2M. Ruvolo3L.A. Turner4S.T. LaBrie51Incyte Genomics, St. Louis, MO, USA1Incyte Genomics, St. Louis, MO, USA1Incyte Genomics, St. Louis, MO, USA1Incyte Genomics, St. Louis, MO, USA1Incyte Genomics, St. Louis, MO, USA1Incyte Genomics, St. Louis, MO, USADNA macroarrays are used in many areas of molecular biology research for applications ranging from gene discovery to gene expression profiling. As an increasing number of specialized macroarrays containing genes related by function or pathway are becoming available, a question that needs to be addressed is the level of hybridization signal specificity between highly similar genes that can be achieved. We have examined the ability of our LifeGrid™ macroarrays to distinguish hybridization signals between closely related genes. We determined the level of cross-hybridization among genes ranging from 52% to 94% sequence identity. Fragments of genes from five protein families were arrayed onto nylon filters. The filters were subsequently hybridized with a 33P-labeled probe prepared from a pool of synthetic mRNA transcripts containing a representative of each protein family. We found that fragments containing sequences with up to 94% sequence identity displayed relatively little cross-hybridization. We conclude that this macroarray system is very specific and that hybridization signals from closely related genes can be reliably measured.https://www.future-science.com/doi/10.2144/02323pf01
spellingShingle N.A. Miller
Q. Gong
R. Bryan
M. Ruvolo
L.A. Turner
S.T. LaBrie
Cross-Hybridization of Closely Related Genes on High-Density Macroarrays
BioTechniques
title Cross-Hybridization of Closely Related Genes on High-Density Macroarrays
title_full Cross-Hybridization of Closely Related Genes on High-Density Macroarrays
title_fullStr Cross-Hybridization of Closely Related Genes on High-Density Macroarrays
title_full_unstemmed Cross-Hybridization of Closely Related Genes on High-Density Macroarrays
title_short Cross-Hybridization of Closely Related Genes on High-Density Macroarrays
title_sort cross hybridization of closely related genes on high density macroarrays
url https://www.future-science.com/doi/10.2144/02323pf01
work_keys_str_mv AT namiller crosshybridizationofcloselyrelatedgenesonhighdensitymacroarrays
AT qgong crosshybridizationofcloselyrelatedgenesonhighdensitymacroarrays
AT rbryan crosshybridizationofcloselyrelatedgenesonhighdensitymacroarrays
AT mruvolo crosshybridizationofcloselyrelatedgenesonhighdensitymacroarrays
AT laturner crosshybridizationofcloselyrelatedgenesonhighdensitymacroarrays
AT stlabrie crosshybridizationofcloselyrelatedgenesonhighdensitymacroarrays