Abstract 2942: Robust sample recovery post-digital PCR for downstream genomic applications

Abstract

Abstract Droplet microfluidics technologies, and particularly, digital PCR have provided highly precise and sensitive detection of genetic targets. There is a strong interest for researchers to reuse sample due to sample scarcity, as well as, to further characterize or validate their experimental findings. Historically, recovery of PCR products for downstream genomic testing applications has been challenging due to the stability of the sample post-reaction. Stilla Technologies has developed Crystal Digital PCR™ and the naica® system, a flexible and highplex droplet and imaging-based digital PCR platform that allows for multi-color detection of genetic targets with the ability to also recover samples after the reaction is performed. This study shows how the 6-color naica digital PCR system workflow can be combined with a simple protocol for sample recovery. DNA contained in the recovered emulsion is extracted using a standard chloroform protocol that takes approximately 1 hour. DNA recovery rate assessment showed high percentage recovery of droplet sample and on average, 70% of the total DNA present from the initial sample recovered. Recovered DNA from the droplet crystal was sequenced and the results were confirmed by qPCR. The droplet and sample recovery protocol post-digital PCR has potential applications to support validation of Crystal Digital PCR results, concordance alongside a multitude of genomic platforms, and further characterization of biomarkers and mutations with profiling platforms such as NGS. Citation Format: Kimberley Gutierrez, Benjamin Foret, Cécile Jovelet, Allison C. Mallory. Robust sample recovery post-digital PCR for downstream genomic applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2942.