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

被引:13
|
作者
Stangegaard, Michael [1 ]
Froslev, Tobias G. [1 ]
Frank-Hansen, Rune [1 ]
Hansen, Anders J. [1 ]
Morling, Niels [1 ]
机构
[1] Univ Copenhagen, Dept Forens Med, Sect Forens Genet, Fac Hlth Sci, DK-2100 Copenhagen, Denmark
来源
JALA | 2011年 / 16卷 / 02期
关键词
forensic genetics; laboratory automation; nucleic acid extraction; magnetic beads; reference samples; NUCLEIC-ACID EXTRACTION;
D O I
10.1016/j.jala.2010.11.003
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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: 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 3.5 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)
引用
收藏
页码:134 / 140
页数:7
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