Gene expression analysis enables valuable insights beyond the static information of a genome sequence by allowing researchers to detect and quantify the expression levels of multiple coding transcripts between samples using RNA sequencing (RNA-Seq). An understanding of differential gene expression between biological or experimental conditions opens the door to identifying the molecular basis of phenotypic differences.
Approaches and Techniques
Gene expression profiling typically involves the isolation or capture of transcribed RNA within a sample, followed by cDNA synthesis and library construction and amplification prior to detection and quantitation by RNA-Seq. Alternatively, total RNA input is converted into a cDNA library, followed by hybridization capture of targets of interest.
The KAPA mRNA HyperPrep Kit provides a streamlined, workflow for the preparation of RNA-Seq libraries containing coding transcripts only. Using oligo-dT beads, polyadenylated transcripts are selectively captured from total RNA samples, after which cDNA is synthesized and libraries constructed for Illumina sequencing. Since RNA enrichment prior to library construction relies on the poly(A) tail of mature, eukaryotic mRNAs, this workflow is only compatible with high-quality RNA inputs from eukaryotic organisms.