High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay

Single nucleotide polymorphisms (SNPs) are the class of DNA variants that are most common in the genome. Technologies are needed that allow relatively highthroughput, high-quality genotyping of SNPs, with rapid assay development for individual SNPs of interest. We have implemented an accurate and hi...

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Main Authors: Phillip A. Morin, Robert Saiz, Atousa Monjazeb
Format: Article
Language:English
Published: Taylor & Francis Group 1999-09-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/99273rr02
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author Phillip A. Morin
Robert Saiz
Atousa Monjazeb
author_facet Phillip A. Morin
Robert Saiz
Atousa Monjazeb
author_sort Phillip A. Morin
collection DOAJ
description Single nucleotide polymorphisms (SNPs) are the class of DNA variants that are most common in the genome. Technologies are needed that allow relatively highthroughput, high-quality genotyping of SNPs, with rapid assay development for individual SNPs of interest. We have implemented an accurate and high-throughput SNP genotyping system using a commercially available fluorescent 5′ exonuclease assay. Optimization of the assay system for low-volume reactions and low-probe concentrations has reduced assay costs by 75%. Using a simple assay optimization process, we successfully developed genotyping assays for 92% of assays attempted (309 out of 335) and have generated over 200 000 genotypes.
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series BioTechniques
spelling doaj-art-a292104d97c74c20a17a7da5b8d4e32f2025-08-20T02:26:02ZengTaylor & Francis GroupBioTechniques0736-62051940-98181999-09-0127353855210.2144/99273rr02High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease AssayPhillip A. Morin0Robert Saiz1Atousa Monjazeb21Axys Pharmaceuticals, La Jolla, CA, USA1Axys Pharmaceuticals, La Jolla, CA, USA1Axys Pharmaceuticals, La Jolla, CA, USASingle nucleotide polymorphisms (SNPs) are the class of DNA variants that are most common in the genome. Technologies are needed that allow relatively highthroughput, high-quality genotyping of SNPs, with rapid assay development for individual SNPs of interest. We have implemented an accurate and high-throughput SNP genotyping system using a commercially available fluorescent 5′ exonuclease assay. Optimization of the assay system for low-volume reactions and low-probe concentrations has reduced assay costs by 75%. Using a simple assay optimization process, we successfully developed genotyping assays for 92% of assays attempted (309 out of 335) and have generated over 200 000 genotypes.https://www.future-science.com/doi/10.2144/99273rr02
spellingShingle Phillip A. Morin
Robert Saiz
Atousa Monjazeb
High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
BioTechniques
title High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
title_full High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
title_fullStr High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
title_full_unstemmed High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
title_short High-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
title_sort high throughput single nucleotide polymorphism genotyping by fluorescent 5 exonuclease assay
url https://www.future-science.com/doi/10.2144/99273rr02
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AT robertsaiz highthroughputsinglenucleotidepolymorphismgenotypingbyfluorescent5exonucleaseassay
AT atousamonjazeb highthroughputsinglenucleotidepolymorphismgenotypingbyfluorescent5exonucleaseassay