Optimized Covaris truXTRAC® FFPE RNA Protocol for Isolating RNA from Laser-Capture Microdissected (LCM) FFPE Tissue for Next Generation Sequencing
Using an adapted Covaris truXTRAC FFPE RNA workflow, it has been shown that total RNA yields could be increased by 8 to 12-fold compared to classical protease-based workflows. In addition, this approach is qualitatively equal, if not superior, as RT-qPCR Cq values were on average 2.3-fold lower. More importantly RNA extracted by this new approach performs well in NGS, a feat previously challenging when using low input RNA. The unique combination of Covaris truXTRAC FFPE RNA kits and Focused-ultrasonicators enables NGS-grade nucleic acids amenable to RNA sequencing, a feat previously challenging when using low input FFPE tissue such LCM-FFPE samples. This workflow is easily transferable to other species, including Human, and to different pathologies, providing a solution to the difficulties previously seen when performing RNA expression studies from FFPE samples.