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Extraction efficiency experiments were conducted to evaluate the percentage of DNA recovered through three extraction methods: a manual phenol-chloroform method, the use of the Qiagen EZ1 Advanced XL Extraction robot, and with Qiagen QIAamp DNA Mini extraction kit. Three DNA sources (extracted DNA, cells, blood) were tested at varying known concentrations. Extracted samples were quantified with the use of droplet digital PCR (ddPCR) with nuclear DNA assays designed and optimized in-house. Droplet digital PCR allows for the absolute quantitation of DNA without the need of a standard curve or calibrant. [ABSTRACT FROM AUTHOR]

In response to the COVID-19 pandemic, the National Institute of Standards and Technology released a synthetic RNA material for SARS-CoV-2 in June 2020. The goal was to rapidly produce a material to support molecular diagnostic testing applications. This material, referred to as Research Grade Test Material 10169, was shipped free of charge to laboratories across the globe to provide a non-hazardous material for assay development and assay calibration. The material consisted of two unique regions of the SARS-CoV-2 genome approximately 4 kb nucleotides in length. The concentration of each synthetic fragment was measured using RT-dPCR methods and confirmed to be compatible with RT-qPCR methods. In this report, the preparation, stability, and limitations of this material are described. [ABSTRACT FROM AUTHOR]