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High Efficiency Hydrodynamic DNA Fragmentation in a Bubbling System

机译:鼓泡系统中的高效流体动力学DNA片段化

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摘要

DNA fragmentation down to a precise fragment size is important for biomedical applications, disease determination, gene therapy and shotgun sequencing. In this work, a cheap, easy to operate and high efficiency DNA fragmentation method is demonstrated based on hydrodynamic shearing in a bubbling system. We expect that hydrodynamic forces generated during the bubbling process shear the DNA molecules, extending and breaking them at the points where shearing forces are larger than the strength of the phosphate backbone. Factors of applied pressure, bubbling time and temperature have been investigated. Genomic DNA could be fragmented down to controllable 1–10 Kbp fragment lengths with a yield of 75.30–91.60%. We demonstrate that the ends of the genomic DNAs generated from hydrodynamic shearing can be ligated by T4 ligase and the fragmented DNAs can be used as templates for polymerase chain reaction. Therefore, in the bubbling system, DNAs could be hydrodynamically sheared to achieve smaller pieces in dsDNAs available for further processes. It could potentially serve as a DNA sample pretreatment technique in the future.
机译:精确到片段大小的DNA片段对于生物医学应用,疾病确定,基因治疗和shot弹枪测序很重要。在这项工作中,基于鼓泡系统中的流体动力剪切,证明了一种廉价,易于操作且高效的DNA片段化方法。我们预计,在起泡过程中产生的流体动力会剪切DNA分子,并在剪切力大于磷酸盐骨架强度的点上延伸和破坏它们。已经研究了施加压力,起泡时间和温度的因素。基因组DNA可以片段化为可控制的1–10 Kbp片段长度,产率为75.30–91.60%。我们证明了由水力剪切产生的基因组DNA的末端可以通过T4连接酶连接,并且片段化的DNA可以用作聚合酶链反应的模板。因此,在起泡系统中,可以对DNA进行流体动力剪切,以使其在dsDNA中获得较小的片段,以供进一步处理。将来有可能用作DNA样品的预处理技术。

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