TY - JOUR
T1 - Automating direct-to-PCR for disaster victim identification
AU - Watherston, J.
AU - Bruce, D.
AU - Ward, J.
AU - Gahan, M. E.
AU - McNevin, D.
PY - 2019
Y1 - 2019
N2 - Direct-to-PCR methodology adds samples directly to PCR tubes offering gains in efficiency and sensitivity. The approach has been applied to a variety of biological sources including blood, saliva, tissue, hair and nail. We added various preservative solutions to a range of biological samples to leech DNA into solution, whilst preserving at room temperature. Tubes containing ‘free DNA’ then followed automated workflows for amplification and capillary electrophoresis. Routine FASS-automated workflows (including DNA extraction and quantification) were compared with published direct-to-PCR methodology and automated amplification of an aliquot of preservative solution. Applying preservative solutions to ~30-year-old blood stains stored at room temperature resulted in recovery of a larger quantity of DNA and more alleles (using PowerPlex 21) when compared with routine automated typing. Trials were extended to blood, saliva, hair and nail, mimicking ante-mortem samples collected in a disaster victim identification effort. Despite slightly lower allelic recovery, the faster processing times, lower costs and storage potential offers advantages for the processing of ante-mortem samples.
AB - Direct-to-PCR methodology adds samples directly to PCR tubes offering gains in efficiency and sensitivity. The approach has been applied to a variety of biological sources including blood, saliva, tissue, hair and nail. We added various preservative solutions to a range of biological samples to leech DNA into solution, whilst preserving at room temperature. Tubes containing ‘free DNA’ then followed automated workflows for amplification and capillary electrophoresis. Routine FASS-automated workflows (including DNA extraction and quantification) were compared with published direct-to-PCR methodology and automated amplification of an aliquot of preservative solution. Applying preservative solutions to ~30-year-old blood stains stored at room temperature resulted in recovery of a larger quantity of DNA and more alleles (using PowerPlex 21) when compared with routine automated typing. Trials were extended to blood, saliva, hair and nail, mimicking ante-mortem samples collected in a disaster victim identification effort. Despite slightly lower allelic recovery, the faster processing times, lower costs and storage potential offers advantages for the processing of ante-mortem samples.
UR - https://hdl.handle.net/1959.7/uws:61075
U2 - 10.1080/00450618.2019.1569145
DO - 10.1080/00450618.2019.1569145
M3 - Article
SN - 0045-0618
VL - 51
SP - S39-S43
JO - Australian Journal of Forensic Sciences
JF - Australian Journal of Forensic Sciences
IS - Suppl. 1
ER -