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Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology
BACKGROUND: The arrival of RNA-seq as a high-throughput method competitive to the established microarray technologies has necessarily driven a need for comparative evaluation. To date, cross-platform comparisons of these technologies have been relatively few in number of platforms analyzed and were...
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/PMC4575490/ https://www.ncbi.nlm.nih.gov/pubmed/26385698 http://dx.doi.org/10.1186/s12864-015-1913-6 |
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author | Yu, Jinsheng Cliften, Paul F. Juehne, Twyla I. Sinnwell, Toni M. Sawyer, Chris S. Sharma, Mala Lutz, Andrew Tycksen, Eric Johnson, Mark R. Minton, Matthew R. Klotz, Elliott T. Schriefer, Andrew E. Yang, Wei Heinz, Michael E. Crosby, Seth D. Head, Richard D. |
author_facet | Yu, Jinsheng Cliften, Paul F. Juehne, Twyla I. Sinnwell, Toni M. Sawyer, Chris S. Sharma, Mala Lutz, Andrew Tycksen, Eric Johnson, Mark R. Minton, Matthew R. Klotz, Elliott T. Schriefer, Andrew E. Yang, Wei Heinz, Michael E. Crosby, Seth D. Head, Richard D. |
author_sort | Yu, Jinsheng |
collection | PubMed |
description | BACKGROUND: The arrival of RNA-seq as a high-throughput method competitive to the established microarray technologies has necessarily driven a need for comparative evaluation. To date, cross-platform comparisons of these technologies have been relatively few in number of platforms analyzed and were typically gene name annotation oriented. Here, we present a more extensive and yet precise assessment to elucidate differences and similarities in performance of numerous aspects including dynamic range, fidelity of raw signal and fold-change with sample titration, and concordance with qRT-PCR (TaqMan). To ensure that these results were not confounded by incompatible comparisons, we introduce the concept of probe mapping directed “transcript pattern”. A transcript pattern identifies probe(set)s across platforms that target a common set of transcripts for a specific gene. Thus, three levels of data were examined: entire data sets, data derived from a subset of 15,442 RefSeq genes common across platforms, and data derived from the transcript pattern defined subset of 7,034 RefSeq genes. RESULTS: In general, there were substantial core similarities between all 6 platforms evaluated; but, to varying degrees, the two RNA-seq protocols outperformed three of the four microarray platforms in most categories. Notably, a fourth microarray platform, Agilent with a modified protocol, was comparable, or marginally superior, to the RNA-seq protocols within these same assessments, especially in regards to fold-change evaluation. Furthermore, these 3 platforms (Agilent and two RNA-seq methods) demonstrated over 80 % fold-change concordance with the gold standard qRT-PCR (TaqMan). CONCLUSIONS: This study suggests that microarrays can perform on nearly equal footing with RNA-seq, in certain key features, specifically when the dynamic range is comparable. Furthermore, the concept of a transcript pattern has been introduced that may minimize potential confounding factors of multi-platform comparison and may be useful for similar evaluations. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1913-6) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4575490 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-45754902015-09-20 Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology Yu, Jinsheng Cliften, Paul F. Juehne, Twyla I. Sinnwell, Toni M. Sawyer, Chris S. Sharma, Mala Lutz, Andrew Tycksen, Eric Johnson, Mark R. Minton, Matthew R. Klotz, Elliott T. Schriefer, Andrew E. Yang, Wei Heinz, Michael E. Crosby, Seth D. Head, Richard D. BMC Genomics Methodology Article BACKGROUND: The arrival of RNA-seq as a high-throughput method competitive to the established microarray technologies has necessarily driven a need for comparative evaluation. To date, cross-platform comparisons of these technologies have been relatively few in number of platforms analyzed and were typically gene name annotation oriented. Here, we present a more extensive and yet precise assessment to elucidate differences and similarities in performance of numerous aspects including dynamic range, fidelity of raw signal and fold-change with sample titration, and concordance with qRT-PCR (TaqMan). To ensure that these results were not confounded by incompatible comparisons, we introduce the concept of probe mapping directed “transcript pattern”. A transcript pattern identifies probe(set)s across platforms that target a common set of transcripts for a specific gene. Thus, three levels of data were examined: entire data sets, data derived from a subset of 15,442 RefSeq genes common across platforms, and data derived from the transcript pattern defined subset of 7,034 RefSeq genes. RESULTS: In general, there were substantial core similarities between all 6 platforms evaluated; but, to varying degrees, the two RNA-seq protocols outperformed three of the four microarray platforms in most categories. Notably, a fourth microarray platform, Agilent with a modified protocol, was comparable, or marginally superior, to the RNA-seq protocols within these same assessments, especially in regards to fold-change evaluation. Furthermore, these 3 platforms (Agilent and two RNA-seq methods) demonstrated over 80 % fold-change concordance with the gold standard qRT-PCR (TaqMan). CONCLUSIONS: This study suggests that microarrays can perform on nearly equal footing with RNA-seq, in certain key features, specifically when the dynamic range is comparable. Furthermore, the concept of a transcript pattern has been introduced that may minimize potential confounding factors of multi-platform comparison and may be useful for similar evaluations. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1913-6) contains supplementary material, which is available to authorized users. BioMed Central 2015-09-18 /pmc/articles/PMC4575490/ /pubmed/26385698 http://dx.doi.org/10.1186/s12864-015-1913-6 Text en © Yu et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Yu, Jinsheng Cliften, Paul F. Juehne, Twyla I. Sinnwell, Toni M. Sawyer, Chris S. Sharma, Mala Lutz, Andrew Tycksen, Eric Johnson, Mark R. Minton, Matthew R. Klotz, Elliott T. Schriefer, Andrew E. Yang, Wei Heinz, Michael E. Crosby, Seth D. Head, Richard D. Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
title | Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
title_full | Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
title_fullStr | Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
title_full_unstemmed | Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
title_short | Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
title_sort | multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575490/ https://www.ncbi.nlm.nih.gov/pubmed/26385698 http://dx.doi.org/10.1186/s12864-015-1913-6 |
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