An instrument for automated purification of nucleic acids from contaminated forensic samples.
An instrument for automated purification of nucleic acids from contaminated forensic samples.
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用于从受污染的法医样品中自动纯化核酸的仪器。
DOI：
10.1016/j.jala.2007.10.008
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Marziali,Andre
中科院分区：
文献类型：
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作者：
Broemeling,DavidJ;Pel,Joel;Gunn,DylanC;Mai,Laura;Thompson,JasonD;Poon,Hiron;Marziali,Andre
法医犯罪现场样品分析，就其性质而言，通常处理其中存在少量核酸的样品，其底物通常导致抑制随后的酶促反应，例如用于短串联重复序列（STR）分析的PCR扩增。常见的基质包括蓝色牛仔裤的粗斜纹棉布，其产生靛蓝染料作为PCR抑制剂，以及土壤，其产生腐殖物质作为抑制剂。这些抑制剂经常与标准柱或基于珠的制备物中的核酸共提取，导致STR分析经常失败。我们提出了一种用于法医样品的DNA纯化的新型仪器，该仪器能够从土壤颗粒、织物和包括固体成分在内的其他复杂样品中高效浓缩核酸。这种新的浓缩过程被称为同步阻力系数改变，它对长电荷聚合物（如DNA）具有固有的选择性，因此能够有效地拒绝已知的污染物。我们提出了一个自动化的样品制备仪器的基础上，这一过程，并根据模拟法医样品的初步结果。
Forensic crime scene sample analysis, by its nature, often deals with samples in which there are low amounts of nucleic acids, on substrates that often lead to inhibition of subsequent enzymatic reactions such as PCR amplification for Short Tandem Repeat (STR) profiling. Common substrates include denim from blue jeans, which yields indigo dye as a PCR inhibitor, and soil, which yields humic substances as inhibitors. These inhibitors frequently co-extract with nucleic acids in standard column or bead-based preps, leading to frequent failure of STR profiling. We present a novel instrument for DNA purification of forensic samples that is capable of highly effective concentration of nucleic acids from soil particulates, fabric, and other complex samples including solid components. The novel concentration process, known as Synchronous Coefficient of Drag Alteration, is inherently selective for long-charged polymers such as DNA, and therefore is able to effectively reject known contaminants. We present an automated sample preparation instrument based on this process, and preliminary results based on mock forensic samples.