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Automated extraction of DNA from blood and PCR setup using a Tecan Freedom EVO liquid handler for forensic genetic STR typing of reference samples

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Michael Stangegaard, Tobias G Frøslev, Rune Frank-Hansen, Anders J Hansen, Niels Morling

We have implemented and validated automated protocols for DNA extraction and PCR setup using a Tecan Freedom EVO liquid handler mounted with the Te-MagS magnetic separation device (Tecan, Männedorf, Switzerland). The protocols were validated for accredited forensic genetic work according to ISO 17025 using the Qiagen MagAttract DNA Mini M48 kit (Qiagen GmbH, Hilden, Germany) from fresh whole blood and blood from deceased individuals. The workflow was simplified by returning the DNA extracts to the original tubes minimizing the risk of misplacing samples. The tubes that originally contained the samples were washed with MilliQ water before the return of the DNA extracts. The PCR was setup in 96-well microtiter plates. The methods were validated for the kits: AmpFlSTR Identifiler, SGM Plus and Yfiler (Applied Biosystems, Foster City, CA), GenePrint FFFL and PowerPlex Y (Promega, Madison, WI). The automated protocols allowed for extraction and addition of PCR master mix of 96 samples within 3.5h. In conclusion, we demonstrated that (1) DNA extraction with magnetic beads and (2) PCR setup for accredited, forensic genetic short tandem repeat typing can be implemented on a simple automated liquid handler leading to the reduction of manual work, and increased quality and throughput.
OriginalsprogEngelsk
TidsskriftJournal of laboratory automation
Vol/bind16
Udgave nummer2
Sider (fra-til)134-40
Antal sider7
ISSN2472-6303
DOI
StatusUdgivet - 1 apr. 2011

Bibliografisk note

Journal of Laboratory Automation 2211-0682 Formerly (until 2011) Association for Laboratory Automation. Journal, The 1535-5535

ID: 33883283