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High Throughput Automated Allele Frequency Estimation by Pyrosequencing
Pyrosequencing is a DNA sequencing method based on the principle of sequencing-by-synthesis and pyrophosphate detection through a series of enzymatic reactions. This bioluminometric, real-time DNA sequencing technique offers unique applications that are cost-effective and user-friendly. In this stud...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2442187/ https://www.ncbi.nlm.nih.gov/pubmed/18628978 http://dx.doi.org/10.1371/journal.pone.0002693 |
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author | Doostzadeh, Julie Shokralla, Shadi Absalan, Farnaz Jalili, Roxana Mohandessi, Sharareh Langston, James W. Davis, Ronald W. Ronaghi, Mostafa Gharizadeh, Baback |
author_facet | Doostzadeh, Julie Shokralla, Shadi Absalan, Farnaz Jalili, Roxana Mohandessi, Sharareh Langston, James W. Davis, Ronald W. Ronaghi, Mostafa Gharizadeh, Baback |
author_sort | Doostzadeh, Julie |
collection | PubMed |
description | Pyrosequencing is a DNA sequencing method based on the principle of sequencing-by-synthesis and pyrophosphate detection through a series of enzymatic reactions. This bioluminometric, real-time DNA sequencing technique offers unique applications that are cost-effective and user-friendly. In this study, we have combined a number of methods to develop an accurate, robust and cost efficient method to determine allele frequencies in large populations for association studies. The assay offers the advantage of minimal systemic sampling errors, uses a general biotin amplification approach, and replaces dTTP for dATP-apha-thio to avoid non-uniform higher peaks in order to increase accuracy. We demonstrate that this newly developed assay is a robust, cost-effective, accurate and reproducible approach for large-scale genotyping of DNA pools. We also discuss potential improvements of the software for more accurate allele frequency analysis. |
format | Text |
id | pubmed-2442187 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-24421872008-07-16 High Throughput Automated Allele Frequency Estimation by Pyrosequencing Doostzadeh, Julie Shokralla, Shadi Absalan, Farnaz Jalili, Roxana Mohandessi, Sharareh Langston, James W. Davis, Ronald W. Ronaghi, Mostafa Gharizadeh, Baback PLoS One Research Article Pyrosequencing is a DNA sequencing method based on the principle of sequencing-by-synthesis and pyrophosphate detection through a series of enzymatic reactions. This bioluminometric, real-time DNA sequencing technique offers unique applications that are cost-effective and user-friendly. In this study, we have combined a number of methods to develop an accurate, robust and cost efficient method to determine allele frequencies in large populations for association studies. The assay offers the advantage of minimal systemic sampling errors, uses a general biotin amplification approach, and replaces dTTP for dATP-apha-thio to avoid non-uniform higher peaks in order to increase accuracy. We demonstrate that this newly developed assay is a robust, cost-effective, accurate and reproducible approach for large-scale genotyping of DNA pools. We also discuss potential improvements of the software for more accurate allele frequency analysis. Public Library of Science 2008-07-16 /pmc/articles/PMC2442187/ /pubmed/18628978 http://dx.doi.org/10.1371/journal.pone.0002693 Text en Doostzadeh et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Doostzadeh, Julie Shokralla, Shadi Absalan, Farnaz Jalili, Roxana Mohandessi, Sharareh Langston, James W. Davis, Ronald W. Ronaghi, Mostafa Gharizadeh, Baback High Throughput Automated Allele Frequency Estimation by Pyrosequencing |
title | High Throughput Automated Allele Frequency Estimation by Pyrosequencing |
title_full | High Throughput Automated Allele Frequency Estimation by Pyrosequencing |
title_fullStr | High Throughput Automated Allele Frequency Estimation by Pyrosequencing |
title_full_unstemmed | High Throughput Automated Allele Frequency Estimation by Pyrosequencing |
title_short | High Throughput Automated Allele Frequency Estimation by Pyrosequencing |
title_sort | high throughput automated allele frequency estimation by pyrosequencing |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2442187/ https://www.ncbi.nlm.nih.gov/pubmed/18628978 http://dx.doi.org/10.1371/journal.pone.0002693 |
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