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Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants
BACKGROUND: Like other structural variants, transposable element insertions can be highly polymorphic across individuals. Their functional impact, however, remains poorly understood. Current genome-wide approaches for genotyping insertion-site polymorphisms based on targeted or whole-genome sequenci...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4494700/ https://www.ncbi.nlm.nih.gov/pubmed/26153459 http://dx.doi.org/10.1186/s12864-015-1700-4 |
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author | Kuhn, Alexandre Ong, Yao Min Quake, Stephen R. Burkholder, William F. |
author_facet | Kuhn, Alexandre Ong, Yao Min Quake, Stephen R. Burkholder, William F. |
author_sort | Kuhn, Alexandre |
collection | PubMed |
description | BACKGROUND: Like other structural variants, transposable element insertions can be highly polymorphic across individuals. Their functional impact, however, remains poorly understood. Current genome-wide approaches for genotyping insertion-site polymorphisms based on targeted or whole-genome sequencing remain very expensive and can lack accuracy, hence new large-scale genotyping methods are needed. RESULTS: We describe a high-throughput method for genotyping transposable element insertions and other types of structural variants that can be assayed by breakpoint PCR. The method relies on next-generation sequencing of multiplex, site-specific PCR amplification products and read count-based genotype calls. We show that this method is flexible, efficient (it does not require rounds of optimization), cost-effective and highly accurate. CONCLUSIONS: This method can benefit a wide range of applications from the routine genotyping of animal and plant populations to the functional study of structural variants in humans. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1700-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4494700 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44947002015-07-08 Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants Kuhn, Alexandre Ong, Yao Min Quake, Stephen R. Burkholder, William F. BMC Genomics Methodology Article BACKGROUND: Like other structural variants, transposable element insertions can be highly polymorphic across individuals. Their functional impact, however, remains poorly understood. Current genome-wide approaches for genotyping insertion-site polymorphisms based on targeted or whole-genome sequencing remain very expensive and can lack accuracy, hence new large-scale genotyping methods are needed. RESULTS: We describe a high-throughput method for genotyping transposable element insertions and other types of structural variants that can be assayed by breakpoint PCR. The method relies on next-generation sequencing of multiplex, site-specific PCR amplification products and read count-based genotype calls. We show that this method is flexible, efficient (it does not require rounds of optimization), cost-effective and highly accurate. CONCLUSIONS: This method can benefit a wide range of applications from the routine genotyping of animal and plant populations to the functional study of structural variants in humans. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1700-4) contains supplementary material, which is available to authorized users. BioMed Central 2015-07-08 /pmc/articles/PMC4494700/ /pubmed/26153459 http://dx.doi.org/10.1186/s12864-015-1700-4 Text en © Kuhn et al. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Kuhn, Alexandre Ong, Yao Min Quake, Stephen R. Burkholder, William F. Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
title | Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
title_full | Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
title_fullStr | Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
title_full_unstemmed | Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
title_short | Read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
title_sort | read count-based method for high-throughput allelic genotyping of transposable elements and structural variants |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4494700/ https://www.ncbi.nlm.nih.gov/pubmed/26153459 http://dx.doi.org/10.1186/s12864-015-1700-4 |
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