Methods and systems for production of dna libraries directly from ffpe tissue

Abstract

Disclosed are methods and compositions for preparing a DNA library from slide-mounted FFPE tissue samples for downstream next-generation sequencing. In one embodiment, a method of preparing the DNA library is disclosed. The method can comprise applying a droplet of a reagent mixture onto a slide-mounted FFPE tissue sample. The reagent mixture can comprise one or more buffer solutions, a cofactor, a nonionic surfactant, a glycerol solution, a gelatin solution, dNTPs, a DNA polymerase, and a primer pool comprising a plurality of forward primers and reverse primers. The method can also comprise stirring the droplet in a circular motion on the slide while scraping portions of the FFPE tissue sample mounted on the slide to yield a reaction mixture. The method can further comprise aspirating the reaction mixture from the slide directly into a pipette tip and dispensing the reaction mixture into a reaction vessel for further amplification.