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

TY - CONF

T1 - Automated extraction of DNA and PCR setup using a Tecan Freedom EVO® liquid handler

AU - Frøslev, Tobias Guldberg

AU - Hansen, Anders Johannes

AU - Stangegaard, Michael

AU - Frank-Hansen, Rune

AU - Morling, Niels

N1 - Titel på proceedings: LabAutomation2009 Sider: 108

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

KW - Faculty of Health and Medical Sciences

M3 - Poster

T2 - LabAutomation2009

Y2 - 25 January 2009 through 28 January 2009

ER -