Estimating the proportion of microarray probes expressed in an RNA sample
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Date
2010
Authors
Shi, W.
de Graaf, C.
Kinkel, S.
Achtman, A.
Baldwin, T.
Schofield, L.
Scott, H.
Hilton, D.
Smyth, G.
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Journal article
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Nucleic Acids Research, 2010; 38(7):2168-2176
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Wei Shi, Carolyn A. de Graaf, Sarah A. Kinkel, Ariel H. Achtman, Tracey Baldwin, Louis Schofield, Hamish S. Scott, Douglas J. Hilton and Gordon K. Smyth
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Abstract
A fundamental question in microarray analysis is the estimation of the number of expressed probes in different RNA samples. Negative control probes available in the latest microarray platforms, such as Illumina whole genome expression BeadChips, provide a unique opportunity to estimate the number of expressed probes without setting a threshold. A novel algorithm was proposed in this study to estimate the number of expressed probes in an RNA sample by utilizing these negative controls to measure background noise. The performance of the algorithm was demonstrated by comparing different generations of Illumina BeadChips, comparing the set of probes targeting well-characterized RefSeq NM transcripts with other probes on the array and comparing pure samples with heterogenous samples. Furthermore, hematopoietic stem cells were found to have a larger transcriptome than progenitor cells. Aire knockout medullary thymic epithelial cells were shown to have significantly less expressed probes than matched wild-type cells.
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© The Author(s) 2010. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.