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Genotyping single-nucleotide polymorphisms by minisequencing using tag arrays
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Molecular Medicine.
2005 (English)In: Microarrays in Clinical Diagnostics / [ed] Thomas O. Joos & Paolo Fortina, Totowa, N.J.: Humana Press, 2005, Vol. 114, 79-92 p.Chapter in book (Other academic)
Abstract [en]

The need for large-scale and high-throughput methods for SNP genotyping has rapidly increased during the last decade. Our system, presented here, combines the highly specific genotyping principle of minisequencing with the advantages of a microarray format that allows highly multiplexed and parallel analysis. Cyclic minisequencing reactions with fluorescently labeled dideoxynucleotides (ddNTPs) are performed in solution using multiplex PCR product as template and detection primers, designed to anneal immediately adjacent and upstream of the SNP site. The detection primers carry unique 5' tag sequences and oligonucleotides complementary to the tag sequence, cTags, are immobilized on a microarray. After extension, the tagged detection primers are allowed to hybridize to the cTags; then the fluorescent signals from the array are measured, and the genotypes are deduced according to the label incorporated. The "array of arrays" format of the system, accomplished by a silicon rubber grid giving separate reaction chambers, allows either 80 or 14 samples to be analyzed for up to 200 or 600 SNPs, respectively, on a single microscope slide.

Place, publisher, year, edition, pages
Totowa, N.J.: Humana Press, 2005. Vol. 114, 79-92 p.
Series
Methods in Molecular Medicine, ISSN 1543-1894 ; 114
Keyword [en]
Genotype, Humans, Oligonucleotide Array Sequence Analysis/instrumentation/methods, Polymorphism; Single Nucleotide, Research Support; Non-U.S. Gov't, Sequence Analysis; DNA/instrumentation/methods
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-76881DOI: 10.1385/1592599230PubMedID: 16156098ISBN: 1588293947 (print)ISBN: 9781588293947 (print)OAI: oai:DiVA.org:uu-76881DiVA: diva2:104793
Available from: 2006-03-09 Created: 2006-03-09 Last updated: 2012-04-25Bibliographically approved

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Syvänen, Ann-Christine

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