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

Abstract

W e 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, M€ annedorf, 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: AmpF[STR 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 3.5 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. ( JALA 2011;16:134-40)

Biomek 3000: The Workhorse in an Automated Accredited Forensic Genetic Laboratory

“…Reference, paternity, and immigration samples were processed in the form of buccal samples on FTA cards (GE Healthcare, Piscataway, NJ), 6 anticoagulated blood or various other types of biological material extracted using a manual Chelex protocol, 7 the M48 extraction kit (Qiagen, Valencia, CA) on a Tecan Freedom EVO (Tecan, Männedorf, Switzerland), 8 a QIAsymphony SP with the QIAsymphony DNA Investigator Kit (Qiagen), or a Qiagen BioRobot EZ1 using the DNA Investigator Kit (Qiagen). For samples processed on the QIAsymphony SP, solubilization of the samples, addition of ATL buffer, and subsequent mixing were accomplished using a Biomek 3000 with a customized instrument deck and an in-house developed method.…”

Approaches to automation of the preanalytical phase of large-scale research in the biobank of the National Medical Research Center for Therapy and Preventive Medicine of the Ministry of Health of Russia

“…Универсальность автоматизированных систем аликвотирования позволяет использовать их в работе с различными типами образцов биологических жидкостей в условиях, когда аликвотирование требует высокой точности [1]. Автоматизированные станции при снабжении их соответствующими модулями могут широко применяться при выделении дезоксирибонуклеиновой кислоты/рибонуклеиновой кислоты (ДНК/РНК), постановке полимеразной цепной реакции (ПЦР), пробоподготовке для геномного секвенирования, проведении иммуноферментного анализа (ИФА), для рассева жидких культур на чашки Петри, а также для выделения мононуклеарных клеток периферической крови [1][2][3][4][5].…”

Validation of the Applied Biosystems RapidHIT ID instrument and ACE GlobalFiler Express sample cartridge

“…However, forensic scientists have continually worked towards reducing the amount of time and labor needed to generate these STR profiles (e.g., updates and developments in automation, direct amplification, polymerase chain reaction (PCR) protocols, etc.) [ 7 – 11 ] as the demand on forensic resources, including trained laboratory personnel, continues to increase while reduced turnaround times are sought. Rapid DNA Systems offer the forensic community a fully automated system capable of processing DNA from a biological sample and interpreting the results in approximately 90 min with minimal human intervention.…”