Bernard Lam's Biography

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Luncheon Technology Spotlight: An Effective NGS Workflow for microRNA Profiling from Liquid Biopsies, Extracellular Vesicles and Small Sample Input - Sample Preservation, RNA Extraction, Amplification and Profiling

Tuesday, 17 November 2015 at 13:00

Add to Calendar ▼2015-11-17 13:00:002015-11-17 14:00:00Europe/LondonLuncheon Technology Spotlight: An Effective NGS Workflow for microRNA Profiling from Liquid Biopsies, Extracellular Vesicles and Small Sample Input - Sample Preservation, RNA Extraction, Amplification and ProfilingSELECTBIOenquiries@selectbiosciences.com

Here, we present an effective workflow for studying miRNAs from various bodily fluids. The workflow involves four important modules - (1) Sample Preservation, (2) RNA Extraction, (3) miRNA Amplification, (4) Detection. Using urine RNA as an example, we demonstrated the importance of preservation as a significant amount of miRNA transcripts could be lost during standard freezing storage. For RNA purification, we demonstrated that the use of silicon carbide-resin technology worked more effectively than traditional phenol:chloroform/silica column-based methods in recovering miRNAs from bodily fluids such as plasma and urine. In particular, the silicon carbide technology does not require the use of carrier RNA and it does not have any bias in GC contents of the miRNAs. Even with effective preservation and RNA extraction, many bodily fluids samples may yield very low amount of RNA (sometimes at the nanogram or picogram range). We will also discuss the methods of quantification of RNA extracted from these liquid biopsies and the expected outcome. Finally, we will present data of a novel procedure that allows amplification of miRNAs from sub-nanogram amount for applications such as RT-qPCR array, microarray and small RNA sequencing.