Royal Canadian Mounted Police, National Services and Research, 1200 Vanier Parkway, Ottawa, Ontario K1G3M8, Canada.
Forensic Sci Int Genet. 2010 Oct;4(5):292-304. doi: 10.1016/j.fsigen.2009.11.001. Epub 2009 Dec 5.
A semi-automated DNA extraction process for casework samples based on the Promega DNA IQ™ system was optimized and validated on TECAN Genesis 150/8 and Freedom EVO robotic liquid handling stations configured with fixed tips and a TECAN TE-Shake™ unit. The use of an orbital shaker during the extraction process promoted efficiency with respect to DNA capture, magnetic bead/DNA complex washes and DNA elution. Validation studies determined the reliability and limitations of this shaker-based process. Reproducibility with regards to DNA yields for the tested robotic workstations proved to be excellent and not significantly different than that offered by the manual phenol/chloroform extraction. DNA extraction of animal:human blood mixtures contaminated with soil demonstrated that a human profile was detectable even in the presence of abundant animal blood. For exhibits containing small amounts of biological material, concordance studies confirmed that DNA yields for this shaker-based extraction process are equivalent or greater to those observed with phenol/chloroform extraction as well as our original validated automated magnetic bead percolation-based extraction process. Our data further supports the increasing use of robotics for the processing of casework samples.
一种基于 Promega DNA IQ™ 系统的半自动化法医样本 DNA 提取方法在 TECAN Genesis 150/8 和 Freedom EVO 机器人液体处理工作站上进行了优化和验证，该工作站配备了固定吸头和 TECAN TE-Shake™ 装置。在提取过程中使用轨道振荡器可提高 DNA 捕获、磁珠/DNA 复合物洗涤和 DNA 洗脱的效率。验证研究确定了这种基于振荡器的方法的可靠性和局限性。对于测试的机器人工作站，关于 DNA 产量的重现性证明非常出色，与手动酚/氯仿提取提供的结果没有显著差异。对污染有土壤的动物：人类血液混合物的 DNA 提取表明，即使存在大量动物血液，也可以检测到人类的特征。对于含有少量生物材料的检材，一致性研究证实，这种基于振荡器的提取方法的 DNA 产量与酚/氯仿提取以及我们原始验证的自动化磁性珠渗滤提取方法相当或更高。我们的数据进一步支持了机器人在法医样本处理中的广泛应用。
