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Gene Expression Assays: RNA-Seq, QPCR and Microarrays

Gene expression assays measure which genes are expressed in different samples, or how the expression of genes change over time or in response to specific treatments. The ATG Shared Resource specializes in using RNA-seq for measuring gene expression.

Next-Gen Sequencing and RNA-Seq

RNA-sequencing (RNA-seq) is a next-generation sequencing approach to measuring gene expression across all the genes at once. Briefly, RNA is isolated, fragmented, converted to cDNA then ligated to adaptors and sequenced. The resulting sequencing "reads" are aligned to the genome. The number of reads aligned to each gene is proportional to the gene expression level. RNA-seq may also provide information about mutations (e.g. SNPs) and alternative RNA splicing, and chromosomal translocations can be detected by analyzing the output for the presence of fusion transcripts that cross the translocation breakpoint. RNA-seq can be successfully performed with high-quality RNA from fresh or frozen tissues or cell lines, and with RNA isolated from pathology samples, such as Formaldehyde-Fixed, Paraffin-Embedded (FFPE) material.

Please contact the ATG Shared Resource staff for more information.