Validation of a first-generation long-oligonucleotide microarray for transcriptional profiling in the pig

• Published in: Genomics (San Diego, Calif.) Vol. 86; no. 5; pp. 618 - 625
• Main Authors: Zhao, Shu-Hong, Recknor, Justin, Lunney, Joan K., Nettleton, Dan, Kuhar, Daniel, Orley, Sarah, Tuggle, Christopher K.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.11.2005; Elsevier
• Subjects: Animals; Biological and medical sciences; complementary DNA; DNA, Complementary; Expressed Sequence Tags; Fundamental and applied biological sciences. Psychology; gene expression; Gene Expression Profiling - methods; Gene Expression Profiling - veterinary; Genes. Genome; Genetics of eukaryotes. Biological and molecular evolution; genomics; In Situ Hybridization; liver; Long oligonucleotide microarray; lungs; microarray technology; Molecular and cellular biology; Molecular genetics; Oligonucleotide Array Sequence Analysis - methods; Oligonucleotide Array Sequence Analysis - veterinary; oligonucleotide microarrays; Pig; Polymerase Chain Reaction; skeletal muscle; small intestine; swine; Swine - genetics; Tissue; Tissue Array Analysis; transcription (genetics); Transcription. Transcription factor. Splicing. Rna processing; Transcriptional profiling; Long oligonucleotide microarray; Tissue; Transcriptional profiling; Pig; Vertebrata; Mammalia; Transcription; Genomics; Oligonucleotide; Artiodactyla; Microarray; Ungulata
• ISSN: 0888-7543; 1089-8646
• DOI: 10.1016/j.ygeno.2005.08.001

A first-generation porcine oligonucleotide set, representing 13,297 cDNAs and ESTs, has been designed by Qiagen–Operon for transcriptional profiling. To validate this set, microarrays containing each 70-mer oligonucleotide, referred to as the Qiagen–NRSP8 array, were hybridized with targets from porcine adult liver, lung, muscle, or small intestine. Transcriptome analyses showed that 11,328 of the oligonucleotides demonstrated expression in at least one tissue. Statistical analyses revealed that 1810 genes showed differential expression among tissues (Bonferroni adjusted p < 0.05). Biological pathways identified by DAVID/EASE analysis using a list of 423 tissue-selective genes matched archetypal pathways in the corresponding human or mouse tissue. Real-time quantitative PCR confirmed expression patterns for 9 of 11 genes tested. Our results demonstrate that this first-generation porcine oligonucleotide array is informative and the specificity is high. This is essential validation for investigators using the Qiagen–NRSP8 array for porcine functional genomics and for using the pig in modeling important physiological problems.