Department of Forensic Science, College of Criminal Justice, Sam Houston State University, 1003 Bowers Blvd., TX, 77341, Huntsville, USA.
Int J Legal Med. 2023 Jul;137(4):949-959. doi: 10.1007/s00414-023-02980-9. Epub 2023 Feb 27.
The InnoXtract™ extraction and purification system is a purification method designed for DNA extraction from low-template samples, specifically rootless hair shafts. Its ability to successfully capture highly fragmented DNA suggests its suitability for use with other challenging sample types, including skeletal remains. However, the lysis and digestion parameters required modifications to successfully optimize the method for this sample type. A two-part digestion was developed utilizing a homebrew digestion buffer (0.5 M EDTA, 0.05% Tween 20, and 100 mM NaCl) and a supplemental lysis with the Hair Digestion Buffer included in the InnoXtract™ kit. Additionally, the magnetic bead volume was modified to improve DNA recovery from these challenging samples. With the altered protocol, the quality and quantity of DNA recovered from InnoXtract™ extracts were comparable to another commercial skeletal extraction method (PrepFiler™ BTA). This modified extraction method successfully purified sufficient amounts of quality DNA from a variety of skeletal samples to produce complete STR profiles. Successful STR typing from surface decomposition, burned, cremated, buried, and embalmed remains indicates the potential of this new method for challenging human identification and missing-person cases.
InnoXtract™ 提取和纯化系统是一种专为从低模板样本（特别是无根毛发）中提取 DNA 而设计的纯化方法。它成功捕获高度碎片化 DNA 的能力表明它适用于其他具有挑战性的样本类型，包括骨骼遗骸。然而，为了成功优化该方法用于这种样本类型，需要对裂解和消化参数进行修改。开发了一种两部分消化方法，利用自制消化缓冲液（0.5 M EDTA、0.05%吐温 20 和 100 mM NaCl）和 InnoXtract™ 试剂盒中包含的头发消化缓冲液进行补充裂解。此外，还修改了磁珠体积，以提高从这些具有挑战性的样本中回收 DNA 的效率。使用修改后的方案，从 InnoXtract™ 提取物中回收的 DNA 的质量和数量与另一种商用骨骼提取方法（PrepFiler™ BTA）相当。这种改进的提取方法成功地从各种骨骼样本中纯化出足够数量的优质 DNA，以产生完整的 STR 图谱。从表面分解、燃烧、火化、埋葬和防腐遗体中成功进行 STR 分型表明，这种新方法在处理具有挑战性的人类识别和失踪人员案件方面具有潜力。
