Investigation of Oxford Nanopore sequencing technologies for forensic purposes

Rebecca Richards presents the feasibility of nanopore sequencing technologies for forensic purposes, using long-range whole mitochondrial genome sequencing as a proof of concept. Three different library preparation methodologies (VolTRAX, Ligation with PCR barcoding and Rapid with PCR Barcoding) were compared using buccal samples and evaluated with respect to the DNA input required, multiplexing capability, ease of use, time, and cost.

The portability, real-time capability, ease of use and low capital cost of the MinION device offers advantages for forensic use in the field. In addition, Flongle, an adapter for the MinION that is more cost-effective for smaller but more frequent testing scenarios, and ‘VolTRAX’, a hand-held microfluidic device that is currently set up to automate the library preparation stage of DNA sequencing.

Following optimisation, the proposed methodology was tested using forensic casework type samples including bloodstains, swabs from bottles and swabs from cans. Finally, an evaluation of Flongle was undertaken to determine if sufficient read depth could be obtained.