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法医学中一种全自动硅胶包被磁珠纯化技术的优化与验证

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

作者信息

Nagy M, Otremba P, Krüger C, Bergner-Greiner S, Anders P, Henske B, Prinz M, Roewer L

机构信息

Institute of Legal Medicine, Charité-University Medicine Berlin, 10115 Berlin, Hannoversche Strasse 6, Germany.

出版信息

Forensic Sci Int. 2005 Aug 11;152(1):13-22. doi: 10.1016/j.forsciint.2005.02.027.

Abstract

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.

摘要

法医DNA分析的自动化程序不仅对高通量样本制备至关重要,而且对于避免常规样本制备过程中的错误也是必需的。基于PCR的法医分析中最关键的阶段是DNA分离,该过程应尽可能多地产生高度纯化的DNA。所使用的提取方法包括污渍和样本的预处理、使用离液剂进行细胞裂解、将DNA与二氧化硅包被的磁性颗粒结合,然后洗脱DNA。我们的工作主要集中在样本制备上,从法医样本中获取尽可能多的生物材料,以及随后的细胞裂解,以创建一个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20000多个常规实验室样本。没有交叉污染的证据。来自少至三个细胞核细胞的DNA可产生可靠的完整STR扩增图谱。DNA在储存两年后仍保持稳定。

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