JAN 27, 2015 2:00 AM PST

Catch-and-Release: Extraction and Purification of NGS-Grade DNA and RNA from FFPE Tissue Samples

Sponsored by: Covaris, Inc., Covaris, Inc.
Speaker
  • Product Manager, Covaris
    Biography
      Dr. Guillaume Durin brings years of experience in protocol and consumables development in the field of sample preparation for molecular biology. Recently, Dr. Durin has worked on developing a DNA and RNA extraction solution for FFPE tissue by utilizing Covaris' patented Adaptive Focused Acoustics Technology and the truXTRAC™ purification kits.

    Abstract
    Formalin Fixed, Paraffin Embedded (FFPE) tissues are rapidly being adopted for transcriptome analysis in conjunction with targeted and whole genome sequencing. Harsh formaldehyde fixation and tissue dehydration of tissue samples results in both a technical challenge to achieve reproducible nucleic acid extraction and molecular analysis, and a workflow challenge in clinical settings. In this webinar, we present truXTRAC™, a highly reproducible method utilizing Covaris Adaptive Focused Acoustics (AFA®) for the extraction and purification of NGS-grade nucleic acids from FFPE tissue.



    Applicable for FFPE cores, sections, and slides, the truXTRAC method utilizes highly controlled and focused acoustic energy for highly efficient paraffin removal and tissue rehydration, which facilitates tissue digestion, crosslink reversal, and nucleic acid release without the use of dangerous organic solvents or inconvenient mineral oils. We will present data from different tissue types and compare the performance of various extraction methods on the same FFPE tissue section for DNA and total RNA. Additionally, the tissuePICK™ device is used to selectively capture regions of interest directly from FFPE tissue slides, enabling an extremely simple workflow with no organic solvents and minimal specialist operator training.



    Importantly, we will present data to illustrate how this method delivers high quality DNA and RNA suitable for NGS library preparation. Our results demonstrate significantly improved DNA and RNA amplificability as indicated by commercial qPCR-based kits designed to assess DNA and RNA quality. Quantitative RT-PCR of beta actin housekeeping gene transcripts of various amplicon lengths for all the tissues types tested indicates a multi-fold increase in yield of amplifiable long transcripts compared to controls isolated with other commercially available FFPE kits.

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