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比较了从 11 种常见干扰物和一个普通拭子中提取萎缩芽孢杆菌孢子 DNA 的 8 种方法。

Comparison of eight methods for the extraction of Bacillus atrophaeus spore DNA from eleven common interferents and a common swab.

机构信息

Detection Department, Defence Science and Technology Laboratory, Porton Down, Salisbury, United Kingdom.

出版信息

PLoS One. 2011;6(7):e22668. doi: 10.1371/journal.pone.0022668. Epub 2011 Jul 26.

Abstract

Eight DNA extraction products or methods (Applied Biosystems PrepFiler Forensic DNA Extraction Kit; Bio-Rad Instagene Only, Bio-Rad Instagene & Spin Column Purification; EpiCentre MasterPure DNA & RNA Kit; FujiFilm QuickGene Mini80; Idaho Technologies 1-2-3 Q-Flow Kit; MoBio UltraClean Microbial DNA Isolation Kit; Sigma Extract-N-Amp Plant and Seed Kit) were adapted to facilitate extraction of DNA under BSL3 containment conditions. DNA was extracted from 12 common interferents or sample types, spiked with spores of Bacillus atropheaus. Resulting extracts were tested by real-time PCR. No one method was the best, in terms of DNA extraction, across all sample types. Statistical analysis indicated that the PrepFiler method was the best method from six dry powders (baking, biological washing, milk, plain flour, filler and talcum) and one solid (Underarm deodorant), the UltraClean method was the best from four liquids (aftershave, cola, nutrient broth, vinegar), and the MasterPure method was the best from the swab sample type. The best overall method, in terms of DNA extraction, across all sample types evaluated was the UltraClean method.

摘要

八种 DNA 提取产品或方法(Applied Biosystems PrepFiler 法医 DNA 提取试剂盒;Bio-Rad Instagene Only、Bio-Rad Instagene 和 Spin Column Purification;EpiCentre MasterPure DNA 和 RNA 试剂盒;FujiFilm QuickGene Mini80;Idaho Technologies 1-2-3 Q-Flow 试剂盒;MoBio UltraClean 微生物 DNA 分离试剂盒;Sigma Extract-N-Amp 植物和种子试剂盒)经过改编,以方便在 BSL3 控制条件下提取 DNA。从 12 种常见干扰物或样品类型中提取 DNA,并在其中添加炭疽芽孢杆菌的孢子。用实时 PCR 测试提取的 DNA。就所有样品类型而言,没有一种方法在 DNA 提取方面是最好的。统计分析表明,在六种干粉(烘烤、生物洗涤、牛奶、普通面粉、填充物和滑石粉)和一种固体(腋下除臭剂)中,PrepFiler 方法是最好的方法;在四种液体(须后水、可乐、营养肉汤、醋)中,UltraClean 方法是最好的方法;在棉签样品类型中,MasterPure 方法是最好的方法。就所有评估的样品类型而言,从所有样品类型中提取 DNA 最好的方法是 UltraClean 方法。

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