Estimation of extraction efficiency by droplet digital PCR.

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]