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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >A novel, integrated forensic microdevice on a rotation‐driven platform: Buccal swab to STR product in less than 2?h
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A novel, integrated forensic microdevice on a rotation‐driven platform: Buccal swab to STR product in less than 2?h

机译：旋转驱动平台上的一种新型，集成的法医微型微型：口腔拭子小于2？H

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This work describes the development of a novel microdevice for forensic DNA processing of reference swabs. This microdevice incorporates an enzyme‐based assay for DNA preparation, which allows for faster processing times and reduced sample handling. Infrared‐mediated PCR (IR‐PCR) is used for STR amplification using a custom reaction mixture, allowing for amplification of STR loci in 45 min while circumventing the limitations of traditional block thermocyclers. Uniquely positioned valves coupled with a simple rotational platform are used to exert fluidic control, eliminating the need for bulky external equipment. All microdevices were fabricated using laser ablation and thermal bonding of PMMA layers. Using this microdevice, the enzyme‐mediated DNA liberation module produced DNA yields similar to or higher than those produced using the traditional (tube‐based) protocol. Initial microdevice IR‐PCR experiments to test the amplification module and reaction (using Phusion Flash/SpeedSTAR) generated near‐full profiles that suffered from interlocus peak imbalance and poor adenylation (significant ?A). However, subsequent attempts using KAPA 2G and Pfu Ultra polymerases generated full STR profiles with improved interlocus balance and the expected adenylated product. A fully integrated run designed to test microfluidic control successfully generated CE‐ready STR amplicons in less than 2?h (1?h of hands‐on time). Using this approach, high‐quality STR profiles were developed that were consistent with those produced using conventional DNA purification and STR amplification methods. This method is a smaller, more elegant solution than current microdevice methods and offers a cheaper, hands‐free, closed‐system alternative to traditional forensic methods.
机译：这项工作描述了用于备注拭子的法医DNA加工的新微型微型微型。该MicroDevice掺入了用于DNA制备的基于酶的测定，其允许更快的加工时间和减少的样品处理。使用定制反应混合物用于红外介导的PCR（IR-PCR）用于STR扩增，允许在45分钟内扩增STR基因座，同时避免传统块热循环仪的局限性。与简单的旋转平台联接的独特定位阀用于施加流体控制，消除了对外部设备的需要。使用PMMA层的激光烧蚀和热粘合来制造所有MicrodeTece。使用该微霉素，酶介导的DNA释放模块产生类似于或高于使用传统（基于管）方案生产的DNA产率。初始MicroDevice IR-PCR实验以测试扩增模块和反应（使用荧光闪光/速度）产生的近乎完全曲线，其患有分隔率不平衡和差的腺苷酸差（重要的α）。然而，随后使用KAPA 2G和PFU超聚合酶的尝试产生了具有改善的跨界平衡和预期的腺苷酸产物的全部STR谱。一种完全集成的运行，设计用于测试微流体控制成功生成的CE-READY STR扩增子，在小于2？H（＆ 1？H的动手时间）。使用这种方法，开发了高质量的STR型材，其与使用常规DNA纯化和STR扩增方法产生的那些一致。该方法是比当前的微型方法更小，更优雅的解决方案，并提供更便宜，免提闭合系统的传统法医方法。

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