Validation of Oligoarrays for Quantitative Exploration of the Transcriptome

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Validation of Oligoarrays for Quantitative Exploration of the Transcriptome

Show simple item record Nygaard, Vigdis Holden, Marit Trimarchi, Jeff Ohno-Machado, Lucila Frigessi, Arnoldo Glad, Ingrid K Wiel, Mark A van de Hovig, Eivind Lyng, Heidi Liu, Fang Kuo, Winston Patrick Cepko, Connie 2011-04-19T21:55:50Z 2008
dc.identifier.citation Nygaard, Vigdis, Fang Liu, Marit Holden, Winston P. Kuo, Jeff Trimarchi, Lucila Ohno-Machado, Connie L. Cepko, et al. 2008. Validation of oligoarrays for quantitative exploration of the transcriptome. BMC Genomics 9: 258. en_US
dc.identifier.issn 1471-2164 en_US
dc.description.abstract Background: Oligoarrays have become an accessible technique for exploring the transcriptome, but it is presently unclear how absolute transcript data from this technique compare to the data achieved with tag-based quantitative techniques, such as massively parallel signature sequencing (MPSS) and serial analysis of gene expression (SAGE). By use of the TransCount method we calculated absolute transcript concentrations from spotted oligoarray intensities, enabling direct comparisons with tag counts obtained with MPSS and SAGE. The tag counts were converted to number of transcripts per cell by assuming that the sum of all transcripts in a single cell was 5·105. Our aim was to investigate whether the less resource demanding and more widespread oligoarray technique could provide data that were correlated to and had the same absolute scale as those obtained with MPSS and SAGE. Results: A number of 1,777 unique transcripts were detected in common for the three technologies and served as the basis for our analyses. The correlations involving the oligoarray data were not weaker than, but, similar to the correlation between the MPSS and SAGE data, both when the entire concentration range was considered and at high concentrations. The data sets were more strongly correlated at high transcript concentrations than at low concentrations. On an absolute scale, the number of transcripts per cell and gene was generally higher based on oligoarrays than on MPSS and SAGE, and ranged from 1.6 to 9,705 for the 1,777 overlapping genes. The MPSS data were on same scale as the SAGE data, ranging from 0.5 to 3,180 (MPSS) and 9 to1,268 (SAGE) transcripts per cell and gene. The sum of all transcripts per cell for these genes was 3.8·105 (oligoarrays), 1.1·105 (MPSS) and 7.6·104 (SAGE), whereas the corresponding sum for all detected transcripts was 1.1·106 (oligoarrays), 2.8·105 (MPSS) and 3.8·105 (SAGE). Conclusion: The oligoarrays and TransCount provide quantitative transcript concentrations that are correlated to MPSS and SAGE data, but, the absolute scale of the measurements differs across the technologies. The discrepancy questions whether the sum of all transcripts within a single cell might be higher than the number of 5·105 suggested in the literature and used to convert tag counts to transcripts per cell. If so, this may explain the apparent higher transcript detection efficiency of the oligoarrays, and has to be clarified before absolute transcript concentrations can be interchanged across the technologies. The ability to obtain transcript concentrations from oligoarrays opens up the possibility of efficient generation of universal transcript databases with low resource demands. en_US
dc.language.iso en_US en_US
dc.publisher BioMed Central en_US
dc.relation.isversionof doi:10.1186/1471-2164-9-258 en_US
dc.relation.hasversion en_US
dash.license LAA
dc.title Validation of Oligoarrays for Quantitative Exploration of the Transcriptome en_US
dc.type Journal Article en_US
dc.description.version Version of Record en_US
dc.relation.journal BMC Genomics en_US Cepko, Connie 2011-04-19T21:55:50Z
dash.affiliation.other HMS^Ophthalmology en_US
dash.affiliation.other HMS^Genetics en_US

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