Abstract

Microarray technologies are established approaches for high throughput gene expression, methylation and genotyping analysis. An accurate mapping of the array probes is essential to generate reliable biological findings. However, manufacturers of the microarray platforms typically provide incomplete and outdated annotation tables, which often rely on older genome and transcriptome versions that differ substantially from up-to-date sequence databases. Here, we present the Re-Annotator, a re-annotation pipeline for microarray probe sequences. It is primarily designed for gene expression microarrays but can also be adapted to other types of microarrays. The Re-Annotator uses a custom-built mRNA reference database to identify the positions of gene expression array probe sequences. We applied Re-Annotator to the Illumina Human-HT12 v4 microarray platform and found that about one quarter (25%) of the probes differed from the manufacturer’s annotation. In further computational experiments on experimental gene expression data, we compared Re-Annotator to another probe re-annotation tool, ReMOAT, and found that Re-Annotator provided an improved re-annotation of microarray probes. A thorough re-annotation of probe information is crucial to any microarray analysis. The Re-Annotator pipeline is freely available at http://sourceforge.net/projects/reannotator along with re-annotated files for Illumina microarrays HumanHT-12 v3/v4 and MouseRef-8 v2.

Highlights

  • Analysis of gene expression profiles under various conditions is one of the corner stones in modern molecular biology research

  • A precise annotation of microarray probe sequences is essential for accurate biological findings and replicability

  • We present a pipeline to re-annotate probe sequences of gene expression microarrays using a custom-built mRNA reference and applied it to three Illumina BeadChip arrays (Human HT-12 v3, v4 and MouseRef-8 v2)

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Summary

Introduction

Analysis of gene expression profiles under various conditions is one of the corner stones in modern molecular biology research. One major challenge in working with gene expression microarrays is the quality of the annotation of the array probes used by the platform. Differences in probe annotations complicate the replication of studies as well as meta-analyses across platforms. The annotations provided by the manufacturers quickly become outdated with every update of the genome assemblies as well as the accompanying annotation tables. The number of annotated transcripts in the RefSeq Gene database

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