title: High-throughput DNA extraction of forensic adhesive tapes.
abstract: Tape-lifting has since its introduction in the early 2000's become a well-established sampling method in forensic DNA analysis. Sampling is quick and straightforward while the following DNA extraction is more challenging due to the "stickiness", rigidity and size of the tape. We have developed, validated and implemented a simple and efficient direct lysis DNA extraction protocol for adhesive tapes that requires limited manual labour. The method uses Chelex beads and is applied with SceneSafe FAST tape. This direct lysis protocol provided higher mean DNA yields than PrepFiler Express BTA on Automate Express, although the differences were not significant when using clothes worn in a controlled fashion as reference material (p=0.13 and p=0.34 for T-shirts and button-down shirts, respectively). Through in-house validation we show that the method is fit-for-purpose for application in casework, as it provides high DNA yields and amplifiability, as well as good reproducibility and DNA extract stability. After implementation in casework, the proportion of extracts with DNA concentrations above 0.01ng/μL increased from 71% to 76%. Apart from providing higher DNA yields compared with the previous method, the introduction of the developed direct lysis protocol also reduced the amount of manual labour by half and doubled the potential throughput for tapes at the laboratory. Generally, simplified manual protocols can serve as a cost-effective alternative to sophisticated automation solutions when the aim is to enable high-throughput DNA extraction of complex crime scene samples.
authors.affiliation: Swedish National Forensic Centre, Linköping, Sweden.
authors.forename: Christina
authors.initials: C
authors.lastname: Forsberg
authors.affiliation: Applied Microbiology, Department of Chemistry, Lund University, Lund, Sweden.
authors.forename: Linda
authors.initials: L
authors.lastname: Jansson
authors.affiliation: Swedish National Forensic Centre, Linköping, Sweden; Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
authors.forename: Ricky
authors.initials: R
authors.lastname: Ansell
authors.affiliation: Swedish National Forensic Centre, Linköping, Sweden; Applied Microbiology, Department of Chemistry, Lund University, Lund, Sweden. Electronic address: johannes.hedman@tmb.lth.se.
authors.forename: Johannes
authors.initials: J
authors.lastname: Hedman
mesh_terms: D000269:Adhesives; D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D018895:Microsatellite Repeats; D011137:Polystyrenes; D011145:Polyvinyls; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000379:methods*
chemical_list: D000269:Adhesives; D011137:Polystyrenes; D011145:Polyvinyls; C006960:chelex; D004247:DNA
keywords: Forensic DNA analysis; Forensic biology; High-throughput; Tape-lifting; Touch DNA; Trace sampling