Improving DNA recovery and sample throughput using the PrepFiler™ Automated Forensic DNA Extraction Kit on two customised Tecan Fluent® 1080 Automated Workstations

One of the primary goals of forensic laboratories performing DNA recovery and profiling is obtaining a high-quality DNA extract from crime scene samples. This is largely dependent on the sensitivity and reliability of the DNA extraction chemistry utilised, as well as the liquid handling and contamination minimisation techniques employed. Automation of DNA extraction methods on large liquid handling platforms allows high-throughput laboratories to apply sensitive chemistries to reliably process a large number of samples, while minimising manual processes and cross-contamination. This study describes the first known implementation of the PrepFiler™ Automated Forensic DNA Extraction Kit on a Tecan Fluent® Gx 1080 Automation Workstation. Two Workstations were customised with the addition of novel “Safe Pipetting Modules” to eliminate sample crossover between wells, which is important in a forensic biology setting to reduce inadvertent DNA transfer. A comparison of DNA extraction efficiency between an optimised PrepFiler™ method and the DNA IQ™ System performed on a Perkin Elmer Janus® Integrator platform showed the optimised PrepFiler™ method consistently extracted a higher yield of DNA from a range of blood inputs, as well as blood and buccal swabs. The PrepFiler™ chemistry also more efficiently removed humic acid and haematin, reducing subsequent PCR inhibition. The subsequent implementation of the optimised PrepFiler™ method onto the Tecan Fluent® Gx workstations showed a further increase in sensitivity, with no evidence of DNA cross-contamination observed. However, the optimised PrepFiler™ method encountered difficulties extracting DNA from fabric substrates, with the PrepFiler Express™ chemistry extracting higher yields on the cartridge-based AutoMate Express™ System. Overall, this study demonstrated the Tecan Fluent® Gx 1080 Automation Workstation is a sensitive, reliable and robust method for DNA extraction using the PrepFiler™ Automated Forensic DNA Extraction Kit, and the addition of the novel Safe Pipetting Module makes this platform an attractive option for forensic biology laboratories where minimising inadvertent DNA transfer is of paramount importance.