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Multi-platform assessment of transcriptional profiling technologies utilizing a precise probe mapping methodology

Authors :
Chris S. Sawyer
Elliott T. Klotz
Jinsheng Yu
Mala Sharma
Richard D. Head
Paul F. Cliften
Eric Tycksen
Wei Yang
Michael E. Heinz
Matthew R Minton
Seth D. Crosby
Mark R. Johnson
Twyla Juehne
Andrew E. Schriefer
Andrew Lutz
Toni M. Sinnwell
Source :
BMC Genomics
Publication Year :
2015
Publisher :
BioMed Central, 2015.

Abstract

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.

Details

Language :
English
ISSN :
14712164
Volume :
16
Issue :
1
Database :
OpenAIRE
Journal :
BMC Genomics
Accession number :
edsair.doi.dedup.....b6a007c4f95c3428e1312accea688997