Conferences \ Circulating Biomarkers World Congress 2015 \ Sample Preparation and Analysis \ Agenda \ Daniela Munafo |
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Increasing Sensitivity of Next Generation Sequencing-based Transcriptome Profiling by Selectively Depleting Abundant RNAsTuesday, 24 March 2015 at 11:00 Add to Calendar ▼2015-03-24 11:00:002015-03-24 12:00:00Europe/LondonIncreasing Sensitivity of Next Generation Sequencing-based Transcriptome Profiling by Selectively Depleting Abundant RNAsSELECTBIOenquiries@selectbiosciences.com Next Generation Sequencing has become the method of choice in research and clinical diagnostics for transcriptome profiling and biomarker discovery. However, whole-transcriptome sequencing is challenging due to the large dynamic range of transcript expression within a total RNA sample. Highly expressed transcripts with minimal biological interest can dominate readouts, masking detection of more informative lower abundant transcripts. I’ll present a method to enrich for RNAs of interest by eliminating unwanted RNAs before sequencing. This method is based on hybridization of probes to the targeted RNA and subsequent enzymatic degradation of the selected RNAs. This method removes abundant RNAs (such us cytoplasmic and mitochondrial ribosomal RNA and hemoglobin transcripts from derivate blood samples) while retaining coding and non-coding RNAs. The reduction of abundant transcripts for RNA-Seq studies significantly increases the ability to detect true biological variations that could not be detected in non-depleted samples. |