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An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences
T cell receptor repertoires can be profiled using next generation sequencing (NGS) to measure and monitor adaptive dynamical changes in response to disease and other perturbations. Genomic DNA-based bulk sequencing is cost-effective but necessitates multiplex target amplification using multiple prim...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10291816/ https://www.ncbi.nlm.nih.gov/pubmed/37365517 http://dx.doi.org/10.1186/s12864-023-09424-z |
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author | Gurun, Burcu Horton, Wesley Murugan, Dhaarini Zhu, Biqing Leyshock, Patrick Kumar, Sushil Byrne, Katelyn T. Vonderheide, Robert H. Margolin, Adam A. Mori, Motomi Spellman, Paul T. Coussens, Lisa M. Speed, Terence P. |
author_facet | Gurun, Burcu Horton, Wesley Murugan, Dhaarini Zhu, Biqing Leyshock, Patrick Kumar, Sushil Byrne, Katelyn T. Vonderheide, Robert H. Margolin, Adam A. Mori, Motomi Spellman, Paul T. Coussens, Lisa M. Speed, Terence P. |
author_sort | Gurun, Burcu |
collection | PubMed |
description | T cell receptor repertoires can be profiled using next generation sequencing (NGS) to measure and monitor adaptive dynamical changes in response to disease and other perturbations. Genomic DNA-based bulk sequencing is cost-effective but necessitates multiplex target amplification using multiple primer pairs with highly variable amplification efficiencies. Here, we utilize an equimolar primer mixture and propose a single statistical normalization step that efficiently corrects for amplification bias post sequencing. Using samples analyzed by both our open protocol and a commercial solution, we show high concordance between bulk clonality metrics. This approach is an inexpensive and open-source alternative to commercial solutions. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-023-09424-z. |
format | Online Article Text |
id | pubmed-10291816 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-102918162023-06-27 An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences Gurun, Burcu Horton, Wesley Murugan, Dhaarini Zhu, Biqing Leyshock, Patrick Kumar, Sushil Byrne, Katelyn T. Vonderheide, Robert H. Margolin, Adam A. Mori, Motomi Spellman, Paul T. Coussens, Lisa M. Speed, Terence P. BMC Genomics Research Article T cell receptor repertoires can be profiled using next generation sequencing (NGS) to measure and monitor adaptive dynamical changes in response to disease and other perturbations. Genomic DNA-based bulk sequencing is cost-effective but necessitates multiplex target amplification using multiple primer pairs with highly variable amplification efficiencies. Here, we utilize an equimolar primer mixture and propose a single statistical normalization step that efficiently corrects for amplification bias post sequencing. Using samples analyzed by both our open protocol and a commercial solution, we show high concordance between bulk clonality metrics. This approach is an inexpensive and open-source alternative to commercial solutions. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-023-09424-z. BioMed Central 2023-06-26 /pmc/articles/PMC10291816/ /pubmed/37365517 http://dx.doi.org/10.1186/s12864-023-09424-z Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Article Gurun, Burcu Horton, Wesley Murugan, Dhaarini Zhu, Biqing Leyshock, Patrick Kumar, Sushil Byrne, Katelyn T. Vonderheide, Robert H. Margolin, Adam A. Mori, Motomi Spellman, Paul T. Coussens, Lisa M. Speed, Terence P. An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences |
title | An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences |
title_full | An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences |
title_fullStr | An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences |
title_full_unstemmed | An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences |
title_short | An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences |
title_sort | open protocol for modeling t cell clonotype repertoires using tcrβ cdr3 sequences |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10291816/ https://www.ncbi.nlm.nih.gov/pubmed/37365517 http://dx.doi.org/10.1186/s12864-023-09424-z |
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