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Optimization and validation of a fully automated silica-coated magnetic beads purification technology in forensics.

机译:在法医中对全自动二氧化硅涂层磁珠纯化技术的优化和验证。

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摘要

Automated procedures for forensic DNA analyses are essential not only for large-throughput sample preparation, but are also needed to avoid errors during routine sample preparation. The most critical stage in PCR-based forensic analysis is DNA isolation, which should yield as much highly purified DNA as possible. The extraction method used consists of pre-treatment of stains and samples, cell lysis using chaotropic reagents, binding of the DNA to silica-coated magnetic particles, followed by elution of the DNA. Our work focuses mainly on sample preparation, obtaining the maximum possible amount of biological material from forensic samples, and the following cell lysis, to create a simple standardized lysis protocol suitable for nearly all forensic material. After optimization and validation, the M-48 BioRobot((R)) workstation has been used for more than 20,000 routine lab samples. There has been no evidence of cross contamination. Resulting DNA from as small as three nuclear cells yield reliable complete STR amplification profiles. The DNA remains stable after 2 years of storage.
机译:法医DNA分析的自动化程序不仅对于大通量样品制备至关重要,而且还需要避免常规样品制备过程中的错误。基于PCR的法医分析中最关键的阶段是DNA分离,它应产生尽可能多的高度纯化的DNA。所用的提取方法包括对污渍和样品进行预处理,使用离液剂进行细胞裂解,将DNA结合到二氧化硅涂层的磁性颗粒上,然后洗脱DNA。我们的工作主要集中在样品制备,从法医样品中获取尽可能多的生物材料以及随后的细胞裂解,以创建适用于几乎所有法医材料的简单标准化裂解方案。经过优化和验证后,M-48 BioRobot(R)工作站已用于20,000多个常规实验室样品。没有证据表明交叉污染。来自三个核细胞的最终DNA产生可靠的完整STR扩增图谱。储存2年后,DNA保持稳定。

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