Forskning ved Københavns Universitet - Københavns Universitet

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Automated extraction of DNA and PCR setup using a Tecan Freedom EVO® liquid handler

Publikation: KonferencebidragPosterForskning

Tobias Guldberg Frøslev, Anders Johannes Hansen, Michael Stangegaard, Rune Frank-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 TeMagS magnetic separation device. The methods were validated for accredited, forensic genetic work according to ISO 17025 using the Qiagen MagAttract® DNA Mini M48 kit from fresh, whole blood and blood from deceased. The methods were simplified by returning the DNA extracts to the original tubes reducing the risk of misplacing samples. The original tubes that had contained the samples were washed with 700 µl Milli-Q water prior to the return of the DNA extracts. The PCR setup protocols were designed for 96 well microtiter plates. The methods were validated for the kits: AmpFlSTR® Identifiler® and Y-filer® (Applied Biosystems), GenePrint® FFFL and PowerPlex® Y (Promega). Within 3.5 hours, 96 samples were extracted and PCR master mix was added. With the Identifiler kit, the number of full DNA profiles was approximately 20% higher with DNA prepared with the robot compared to that obtained with DNA prepared manually with the Chelex method. In conclusion, we demonstrated that (1) DNA extraction with magnetic beads and (2) PCR setup for accredited, forensic genetic DNA typing can be implemented on a simple robot leading to the reduction of manual work as well as increased quality and throughput.
OriginalsprogEngelsk
Publikationsdato2009
Antal sider1
StatusUdgivet - 2009
BegivenhedLabAutomation2009 - Palm Springs 2009, USA
Varighed: 25 jan. 200928 jan. 2009

Konference

KonferenceLabAutomation2009
LandUSA
ByPalm Springs 2009
Periode25/01/200928/01/2009

Bibliografisk note

Titel på proceedings: LabAutomation2009
Sider: 108

ID: 14880627