Validation of automated PCR-setup of the Quantifiler® Trio DNA Quantification kit on the Biomek® 4000 Laboratory Automation Workstation

Abstract The accurate quantification of DNA is critical for choosing the optimal workflow for evidence samples in forensic genetics. Thermo Fisher Scientific has recently released the Quantifiler ® Trio DNA Quantification kit, which they announce has several improvements compared to previous quantification kits, such as increased accuracy in determining the level of total human and male DNA in low-template samples. Since automation of PCR-setups reduces the manual workload and the risk for human errors, we have chosen to validate the PCR-setup of the Quantifiler ® Trio kit on a small benchtop liquid handler, the Biomek ® 4000 (Beckman Coulter). Particularly, we tested the reproducibility of the kit and its performance on low-template samples, focusing on the human and male targets. Furthermore, we conducted a mixture study with different male to female ratios, correlating quantification results to DNA-profiles (NGM Select™) of these mixtures. In respect to automation, we have also examined the level of cross contamination during plate setup. Our results show that the Biomek ® 4000 is as suited as the manual procedure for setup of the Quantifiler ® Trio. No evidence of cross contamination was observed. We found that the automated setup of the Quantifler ® Trio kit on the Biomek ® 4000 results in consistent and reliable quantification results, both for total human DNA and male DNA. Furthermore, the Biomek ® 4000 represents a flexible and user-friendly platform for automated PCR-setups. Implementation of the Quantifiler ® Trio seems to improve the STR-typing workflow for low-template samples and mixtures with a minor male contributor in focus of investigation.