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Optimising the analysis of transcript data using high density oligonucleotide arrays and genomic DNA-based probe selection
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Graham, Neil S., Broadley, Martin R., Hammond, John P., White, Philip J. and May, Sean T. (2007) Optimising the analysis of transcript data using high density oligonucleotide arrays and genomic DNA-based probe selection. BMC Genomics, Vol.8 (No.344). doi:10.1186/1471-2164-8-344
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Official URL: http://dx.doi.org/10.1186/1471-2164-8-344
Abstract
Background: Affymetrix GeneChip arrays are widely used for transcriptomic studies in a diverse range of species. Each gene is represented on a GeneChip array by a probe-set, consisting of up to 16 probe-pairs. Signal intensities across probe-pairs within a probe-set vary in part due to different physical hybridisation characteristics of individual probes with their target labelled transcripts. We
have previously developed a technique to study the transcriptomes of heterologous species based
on hybridising genomic DNA (gDNA) to a GeneChip array designed for a different species, and subsequently using only those probes with good homology.
Results: Here we have investigated the effects of hybridising homologous species gDNA to study the transcriptomes of species for which the arrays have been designed. Genomic DNA from Arabidopsis thaliana and rice (Oryza sativa) were hybridised to the Affymetrix Arabidopsis ATH1 and Rice Genome GeneChip arrays respectively. Probe selection based on gDNA hybridisation
intensity increased the number of genes identified as significantly differentially expressed in two
published studies of Arabidopsis development, and optimised the analysis of technical replicates obtained from pooled samples of RNA from rice.
Conclusion: This mixed physical and bioinformatics approach can be used to optimise estimates of gene expression when using GeneChip arrays.
| Item Type: | Journal Article | ||||
|---|---|---|---|---|---|
| Subjects: | Q Science > QR Microbiology | ||||
| Divisions: | Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010) | ||||
| Library of Congress Subject Headings (LCSH): | DNA analysis, Biological systems, DNA microarrays | ||||
| Journal or Publication Title: | BMC Genomics | ||||
| Publisher: | BioMed Central Ltd. | ||||
| ISSN: | 1471-2164 | ||||
| Official Date: | 1 October 2007 | ||||
| Dates: |
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| Volume: | Vol.8 | ||||
| Number: | No.344 | ||||
| DOI: | 10.1186/1471-2164-8-344 | ||||
| Status: | Peer Reviewed | ||||
| Access rights to Published version: | Open Access | ||||
| Funder: | Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Great Britain. Dept. for Environment, Food & Rural Affairs (DEFRA), Scotland. Environment and Rural Affairs Dept. (SEERAD) | ||||
| Grant number: | HH3501SFV, HH3504SFV (DEFRA) |
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