Evaluation of the Amicon® Ultra-15 centrifugal unit for large volume DNA extraction

Efficient recovery of DNA from large volume forensic samples continues to pose significant challenges for successful DNA analysis due to limitations with upscaling current forensic DNA extraction methods. This study aimed to develop a large-volume DNA extraction protocol incorporating the Amicon® Ultra-15 Centrifugal Unit. Two variations, utilising a wash method and a lysis method, were compared to a traditional centrifugal cell pellet method and a baseline yield. The Amicon® lysis and Amicon® wash methods had higher DNA recovery rates compared to baseline (84% and 43% respectively) than the centrifugal cell pellet method (31%). Whilst the Amicon® lysis method produced the best DNA concentration, a procedural issue was observed where a precipitate formed when Buffer AL was added during the extraction process. These findings suggest the Amicon® lysis method offers the most potential as a large volume DNA extraction method for forensic biology, however, further optimisation is required to identify a suitable lysis buffer that maintains the high DNA recovery whilst minimising precipitate formation.