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Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays
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| Zeitschriftentitel: | Genome Research |
|---|---|
| Personen und Körperschaften: | , , , , |
| In: | Genome Research, 7, 1997, 6, S. 606-614 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Cold Spring Harbor Laboratory
|
| Schlagwörter: |
| author_facet |
Pastinen, Tomi Kurg, Ants Metspalu, Andres Peltonen, Leena Syvänen, Ann-Christine Pastinen, Tomi Kurg, Ants Metspalu, Andres Peltonen, Leena Syvänen, Ann-Christine |
|---|---|
| author |
Pastinen, Tomi Kurg, Ants Metspalu, Andres Peltonen, Leena Syvänen, Ann-Christine |
| spellingShingle |
Pastinen, Tomi Kurg, Ants Metspalu, Andres Peltonen, Leena Syvänen, Ann-Christine Genome Research Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays Genetics (clinical) Genetics |
| author_sort |
pastinen, tomi |
| spelling |
Pastinen, Tomi Kurg, Ants Metspalu, Andres Peltonen, Leena Syvänen, Ann-Christine 1088-9051 1549-5469 Cold Spring Harbor Laboratory Genetics (clinical) Genetics http://dx.doi.org/10.1101/gr.7.6.606 <jats:p>We describe a method for multiplex detection of mutations in which the solid-phase minisequencing principle is applied to an oligonucleotide array format. The mutations are detected by extending immobilized primers that anneal to their template sequences immediately adjacent to the mutant nucleotide positions with single labeled dideoxynucleoside triphosphates using a DNA polymerase. The arrays were prepared by coupling one primer per mutation to be detected on a small glass area. Genomic fragments spanning nine disease mutations, which were selected as targets for the assay, were amplified in multiplex PCR reactions and used as templates for the minisequencing reactions on the primer array. The genotypes of homozygous and heterozygous genomic DNA samples were unequivocally defined at each analyzed nucleotide position by the highly specific primer extension reaction. In a comparison to hybridization with immobilized allele-specific probes in the same assay format, the power of discrimination between homozygous and heterozygous genotypes was one order of magnitude higher using the minisequencing method. Therefore, single-nucleotide primer extension is a promising principle for future high-throughput mutation detection and genotyping using high density DNA-chip technology.</jats:p> Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays Genome Research |
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Cold Spring Harbor Laboratory, 1997 |
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1997 |
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Cold Spring Harbor Laboratory |
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Genome Research |
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| title |
Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_unstemmed |
Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_full |
Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_fullStr |
Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_full_unstemmed |
Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_short |
Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_sort |
minisequencing: a specific tool for dna analysis and diagnostics on oligonucleotide arrays |
| topic |
Genetics (clinical) Genetics |
| url |
http://dx.doi.org/10.1101/gr.7.6.606 |
| publishDate |
1997 |
| physical |
606-614 |
| description |
<jats:p>We describe a method for multiplex detection of mutations in which the solid-phase minisequencing principle is applied to an oligonucleotide array format. The mutations are detected by extending immobilized primers that anneal to their template sequences immediately adjacent to the mutant nucleotide positions with single labeled dideoxynucleoside triphosphates using a DNA polymerase. The arrays were prepared by coupling one primer per mutation to be detected on a small glass area. Genomic fragments spanning nine disease mutations, which were selected as targets for the assay, were amplified in multiplex PCR reactions and used as templates for the minisequencing reactions on the primer array. The genotypes of homozygous and heterozygous genomic DNA samples were unequivocally defined at each analyzed nucleotide position by the highly specific primer extension reaction. In a comparison to hybridization with immobilized allele-specific probes in the same assay format, the power of discrimination between homozygous and heterozygous genotypes was one order of magnitude higher using the minisequencing method. Therefore, single-nucleotide primer extension is a promising principle for future high-throughput mutation detection and genotyping using high density DNA-chip technology.</jats:p> |
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| author | Pastinen, Tomi, Kurg, Ants, Metspalu, Andres, Peltonen, Leena, Syvänen, Ann-Christine |
| author_facet | Pastinen, Tomi, Kurg, Ants, Metspalu, Andres, Peltonen, Leena, Syvänen, Ann-Christine, Pastinen, Tomi, Kurg, Ants, Metspalu, Andres, Peltonen, Leena, Syvänen, Ann-Christine |
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| description | <jats:p>We describe a method for multiplex detection of mutations in which the solid-phase minisequencing principle is applied to an oligonucleotide array format. The mutations are detected by extending immobilized primers that anneal to their template sequences immediately adjacent to the mutant nucleotide positions with single labeled dideoxynucleoside triphosphates using a DNA polymerase. The arrays were prepared by coupling one primer per mutation to be detected on a small glass area. Genomic fragments spanning nine disease mutations, which were selected as targets for the assay, were amplified in multiplex PCR reactions and used as templates for the minisequencing reactions on the primer array. The genotypes of homozygous and heterozygous genomic DNA samples were unequivocally defined at each analyzed nucleotide position by the highly specific primer extension reaction. In a comparison to hybridization with immobilized allele-specific probes in the same assay format, the power of discrimination between homozygous and heterozygous genotypes was one order of magnitude higher using the minisequencing method. Therefore, single-nucleotide primer extension is a promising principle for future high-throughput mutation detection and genotyping using high density DNA-chip technology.</jats:p> |
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| spelling | Pastinen, Tomi Kurg, Ants Metspalu, Andres Peltonen, Leena Syvänen, Ann-Christine 1088-9051 1549-5469 Cold Spring Harbor Laboratory Genetics (clinical) Genetics http://dx.doi.org/10.1101/gr.7.6.606 <jats:p>We describe a method for multiplex detection of mutations in which the solid-phase minisequencing principle is applied to an oligonucleotide array format. The mutations are detected by extending immobilized primers that anneal to their template sequences immediately adjacent to the mutant nucleotide positions with single labeled dideoxynucleoside triphosphates using a DNA polymerase. The arrays were prepared by coupling one primer per mutation to be detected on a small glass area. Genomic fragments spanning nine disease mutations, which were selected as targets for the assay, were amplified in multiplex PCR reactions and used as templates for the minisequencing reactions on the primer array. The genotypes of homozygous and heterozygous genomic DNA samples were unequivocally defined at each analyzed nucleotide position by the highly specific primer extension reaction. In a comparison to hybridization with immobilized allele-specific probes in the same assay format, the power of discrimination between homozygous and heterozygous genotypes was one order of magnitude higher using the minisequencing method. Therefore, single-nucleotide primer extension is a promising principle for future high-throughput mutation detection and genotyping using high density DNA-chip technology.</jats:p> Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays Genome Research |
| spellingShingle | Pastinen, Tomi, Kurg, Ants, Metspalu, Andres, Peltonen, Leena, Syvänen, Ann-Christine, Genome Research, Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays, Genetics (clinical), Genetics |
| title | Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_full | Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_fullStr | Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_full_unstemmed | Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_short | Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| title_sort | minisequencing: a specific tool for dna analysis and diagnostics on oligonucleotide arrays |
| title_unstemmed | Minisequencing: A Specific Tool for DNA Analysis and Diagnostics on Oligonucleotide Arrays |
| topic | Genetics (clinical), Genetics |
| url | http://dx.doi.org/10.1101/gr.7.6.606 |