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The Hows and Whys of Early Steps in RNA Analysis

Leta is a Senior R&D Scientist at Promega Corporation, based in Madison, WI. She received a B.A. in Biology and Physics from Lawrence University, and a PhD in Genetics from Harvard University.  As a graduate student, Leta mapped novel genetic mutations causing muscular dystrophy, using zebrafish as a model organism. Leta has been with Promega for 8 years, focusing on nucleic acid amplification technologies.

  • Leta Steffen, PhD

  • Senior Research Scientist

  • Original Webinar Date: Tuesday, August 11, 2015

Learn about the early steps of purification, protection from degradation and quantitation to improve your downstream analysis.

When you analyze RNA expression levels using methods such as RT-qPCR or RNA-seq, how much thought do you put into the upstream steps of purification and quantitation? Many researchers take these steps for granted, but in fact, the quality and quantity of the input RNA has a dramatic effect on downstream analysis success. RNA, unlike DNA, is also highly susceptible to degradation by RNases present in sample or introduced during processing, and not surprisingly the use of degraded RNA in an experiment reduces the quality of the final results too.

This webinar will provide an overview of the different methods available for purifying RNA from both fresh and FFPE mammalian samples, protecting it from degradation, and assessing its quality prior to analysis with the goal of increasing the success and quality of your downstream analyses. We will describe the various purification chemistries available such as silica- and cellulose-based systems, different throughput options and the pros and cons of each system. RNA assessment methods for RNA quality, quantity, and purity will be addressed. Finally, we will spend some time discussing RNase contamination and the use of RNase inhibitors to protect RNA during purification, handling and downstream analysis.