Extraction and Purification of High-Quality Nucleic Acids from FFPE Samples in an Automated 96-Well Format with truXTRAC FFPE total NA Gen 3 (96)

Covaris has developed comprehensive portfolio of kits that are incorporating Adaptive Focused Acoustics® (AFA) to extract nucleic acids from FFPE (Formalin-fixed Paraffin-embedded) tissue samples.  The truXTRAC FFPE series of extraction and purification kits include DNA- or RNA-only as well total nucleic acid (sequential RNA and DNA from the same sample) extraction, coupled with column or magnetic bead-based purification.   These manual first generation kits are using AFA to deparaffinize, hydrate, homogenize and extract nucleic acids in a single step and are adaptable for semi-automation (e.g., AFA-based extraction coupled with KingFisher Flex and Duo purification).

To support the increasing demand for automated high throughput and full automation, and to maintain the same high AFA-quality of the extracted RNA and DNA first generation chemistry has been modified to accommodate a 96-well format on a liquid handler without manual intervention.  96 individual 0.5 mL Matrix tubes with automated bar-code reader-access at the tube bottom (with automated capping/decapping) can be processed in parallel facilitated by a “Shuttle” adaptor to transfer all 96 tubes to the Covaris R230 (on-deck) Focused-ultrasonicator, the heat-blocks, automated centrifuge, capper/decapper and the magnet stations.  The workflow was also improved with an initial deparaffinization step using a novel non-toxic reagent to eliminate possible problems with automated pipetting steps during supernatant removal.   Subsequent AFA-treatments ensure that any residual wax is actively removed guaranteeing higher yields as well as high quality nucleic acids.  The modification also allowed to both simplify and speed up the magnetic bead-based purification workflow without jeopardizing yield and quality of RNA and DNA fractions.