A Comparison of Control Spleen Gene Expression Using Microarray and High Through-Put Sequencing

Mirabito, Anne
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University of Delaware
Gene expression is the process of transcribing the DNA that resides on a gene into mRNA and translating the mRNA into protein. Studying allows researchers to decipher the functions of genes and their gene products, i.e. proteins. The objective of this project was to compare two methods of monitoring gene expression, microarray technology and Illumina sequencing technology, using the same avian spleen sample. A microarray consists of a glass slide that contains samples of many genes distributed in an ordered pattern. In this case, a 4,949-element avian innate immunity cDNA microarray (AIIM) was used. Illumina sequencing technology is based on massively parallel sequencing of millions of fragments using reversible terminator based sequencing chemistry in conjunction with the Illumina Genome Analyzer. The results obtained using these two methods were compared on a global level, in terms of housekeeping genes, and JAK pathway gene expression. In addition to these comparisons, some practical considerations about the two processes were made. Many of the most highly expressed genes in the Illumina data also showed expression levels with the array, through there was not a clear trend between them. Both of these technologies can be implemented to study gene expression. Though Illumina technology may be more quantitative and possibly more sensitive, the data generated from Illumina sequencing does not yet have a standard quantification method, as does microarray data.