Automated Extraction of DNA from Blood and PCR Setup using a Tecan Freedom EVO Liquid Handler for Forensic Genetic STR Typing of Reference Samples

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, Mannedorf, 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: AmpFSTR Identifier, SGM Plus and Yfiler (Applied Biosystems, Foster City, CA), GenePrint FFFL and PowerPlex Y (Promega, Madison, Wl). The automated protocols allowed for extraction and addition of PCR master mix of 96 samples within 3.5 h. 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.

[1]  Craig Yensen,et al.  Automated Processing of Forensic Casework Samples Using Robotic Workstations Equipped with Nondisposable Tips: Contamination Prevention , 2008, Journal of forensic sciences.

[2]  Z Krnajski,et al.  Performance verification of the Maxwell 16 Instrument and DNA IQ Reference Sample Kit for automated DNA extraction of known reference samples , 2007, Forensic science, medicine, and pathology.

[3]  Birgit Bayer,et al.  Application of the BioRobot EZ1 in a forensic laboratory. , 2005, Legal medicine.

[4]  K. Schmid,et al.  Comparison of manual and automated nucleic acid extraction from whole‐blood samples , 2007, Journal of clinical laboratory analysis.

[5]  N. Morling,et al.  Automated extraction of DNA and PCR setup using a Tecan Freedom EVO® liquid handler , 2009 .

[6]  M Prinz,et al.  Optimization and validation of a fully automated silica-coated magnetic beads purification technology in forensics. , 2005, Forensic science international.

[7]  R Higuchi,et al.  Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. , 2013, BioTechniques.

[8]  Niels Morling,et al.  A Simple Method for Validation and Verification of Pipettes Mounted on Automated Liquid Handlers , 2011, Journal of laboratory automation.

[9]  Christina Valgren,et al.  A comparison of three automated DNA purification methods in Forensic casework , 2008 .

[10]  Ejvor Sassarinis Jansson,et al.  Hand and shoulder ailments among laboratory technicians using modern plunger-operated pipettes. , 1994, Applied ergonomics.

[11]  Chantal J. Frégeau,et al.  Optimized Configuration of Fixed-Tip Robotic Liquid-Handling Stations for the Elimination of Biological Sample Cross-Contamination , 2007 .

[12]  M. Catton,et al.  A comparative evaluation of the sensitivity of two automated and two manual nucleic acid extraction methods for the detection of norovirus by RT-PCR. , 2008, Journal of virological methods.