DNA Shearing Protocols for Ion Torrent Platforms with the M220 Focused-ultrasonicator

Protocol: DNA Shearing Protocols for Ion Torrent Platforms with the M220 Focused-ultrasonicator

Document Number: 010194

PRODUCT INSERT: Loading and Rebuilding Process Vessel SST 2ml (500350) Using Prep-Station (500345)

PRODUCT INSERT:  Loading and Rebuilding Process Vessel SST 2ml (500350) Using Prep-Station (500345)

Protocol: truXTRAC® FFPE total NA Kit – Magnetic Bead Purification (25)

truXTRAC® FFPE total NA Kit – Magnetic Bead Purification (25)

Document Number: 010455

Part Number: 520246

Optimized Covaris truXTRAC® FFPE RNA Protocol for Isolating RNA from Laser-Capture Microdissected (LCM) FFPE Tissue for Next Generation Sequencing

SUMMARY
It is well characterized that RNA extraction from FFPE tissues can be challenging, often producing low yields and/or poor-quality RNA which can adversely affect success of downstream applications such as Next-Generation Sequencing (NGS). In this application note we describe an optimized workflow using Covaris truXTRAC FFPE RNA kits with deparaffinized, stained LCM-FFPE tissues of cancerassociated stroma (CAS) and matched normal stroma derived from canine mammary tumors. Using this adapted truXTRAC workflow, compared to the classical protease-based RNA isolation methods, total RNA yields were increased by 8- to 12-fold and RT-qPCR Cq values were 2.3-fold lower on average, signifying better performance. Finally, this improved quantity and quality of RNA performs favorably well in NGS workflows, overcoming the challenges of using low input RNA in RNA sequencing.