Comprehensive quality control utilizing the prehybridization third-dye image leads to accurate gene expression measurements by cDNA microarrays. BMC Bioinformatics 2006 Aug 14;7:378
Date
08/16/2006Pubmed ID
16907976Pubmed Central ID
PMC1563483DOI
10.1186/1471-2105-7-378Scopus ID
2-s2.0-33748657365 (requires institutional sign-in at Scopus site) 9 CitationsAbstract
BACKGROUND: Gene expression profiling using microarrays has become an important genetic tool. Spotted arrays prepared in academic labs have the advantage of low cost and high design and content flexibility, but are often limited by their susceptibility to quality control (QC) issues. Previously, we have reported a novel 3-color microarray technology that enabled array fabrication QC. In this report we further investigated its advantage in spot-level data QC.
RESULTS: We found that inadequate amount of bound probes available for hybridization led to significant, gene-specific compression in ratio measurements, increased data variability, and printing pin dependent heterogeneities. The impact of such problems can be captured through the definition of quality scores, and efficiently controlled through quality-dependent filtering and normalization. We compared gene expression measurements derived using our data processing pipeline with the known input ratios of spiked in control clones, and with the measurements by quantitative real time RT-PCR. In each case, highly linear relationships (R2 > 0.94) were observed, with modest compression in the microarray measurements (correction factor < 1.17).
CONCLUSION: Our microarray analytical and technical advancements enabled a better dissection of the sources of data variability and hence a more efficient QC. With that highly accurate gene expression measurements can be achieved using the cDNA microarray technology.
Author List
Wang X, Jia S, Meyer L, Xiang B, Chen LY, Jiang N, Moreno C, Jacob HJ, Ghosh S, Hessner MJAuthors
Martin J. Hessner PhD Professor in the Pediatrics department at Medical College of WisconsinShuang Jia Biostatistician III in the Pediatrics department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
AnimalsFluorescent Dyes
Gene Expression Profiling
Liver
Oligonucleotide Array Sequence Analysis
Quality Control
RNA
Rats
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
