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Rapid Nucleic Acid Extraction and Purification Using a Miniature Ultrasonic Technique

机译:使用微型超声波技术快速提取和纯化核酸

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摘要

Miniature ultrasonic lysis for biological sample preparation is a promising technique for efficient and rapid extraction of nucleic acids and proteins from a wide variety of biological sources. Acoustic methods achieve rapid, unbiased, and efficacious disruption of cellular membranes while avoiding the use of harsh chemicals and enzymes, which interfere with detection assays. In this work, a miniature acoustic nucleic acid extraction system is presented. Using a miniature bulk acoustic wave (BAW) transducer array based on 36???° Y-cut lithium niobate, acoustic waves were coupled into disposable laminate-based microfluidic cartridges. To verify the lysing effectiveness, the amount of liberated ATP and the cell viability were measured and compared to untreated samples. The relationship between input power, energy dose, flow-rate, and lysing efficiency were determined. DNA was purified on-chip using three approaches implemented in the cartridges: a silica-based sol-gel silica-bead filled microchannel, nucleic acid binding magnetic beads, and Nafion-coated electrodes. Using E. coli, the lysing dose defined as ATP released per joule was 2.2???? greater, releasing 6.1???? more ATP for the miniature BAW array compared to a bench-top acoustic lysis system. An electric field-based nucleic acid purification approach using Nafion films yielded an extraction efficiency of 69.2% in 10 min for 50 ???μL samples.
机译:用于生物样品制备的微型超声裂解技术是从多种生物来源中高效,快速提取核酸和蛋白质的有前途的技术。声学方法可快速,无偏且有效地破坏细胞膜,同时避免使用会干扰检测分析的刺激性化学物质和酶。在这项工作中,提出了微型声学核酸提取系统。使用基于36°Y切割的铌酸锂的微型体声波(BAW)换能器阵列,将声波耦合到一次性的基于层压材料的微流体弹药筒中。为了验证裂解效果,测量了释放的ATP的量和细胞活力,并与未处理的样品进行了比较。确定了输入功率,能量剂量,流速和裂解效率之间的关系。使用盒中实现的三种方法在芯片上纯化DNA:基于二氧化硅的溶胶凝胶硅胶珠填充微通道,核酸结合磁珠和Nafion涂层电极。使用大肠杆菌,每焦耳释放的ATP的裂解剂量为2.2 ????。更大,发布6.1 ????与台式声裂解系统相比,微型BAW阵列具有更多的ATP。使用Nafion膜的基于电场的核酸纯化方法在50分钟内对50μL样品的提取效率为69.2%。

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