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New Detection Modality for Label-Free Quantification of DNA in Biological Samples via Superparamagnetic Bead Aggregation

机译:通过超顺磁珠聚集对生物样品中的DNA进行无标签定量的新检测方式

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摘要

Combining DNA and superparamagnetic beads in a rotating magnetic field produces multipartide aggregates that are visually striking, enabling label-free optical detection and quantification of DNA at levels in the picogram per microliter range. DNA in biological samples can be quantified directly by simple analysis of optical images of microfluidic wells placed on a magnetic stirrer without prior DNA purification. Aggregation results from DNA/bead interactions driven either by the presence of a chaotrope (a nonspecific trigger for aggregation) or by hybridization with oligonudeo-tides on functionalized beads (sequence-specific). This paper demonstrates quantification of DNA with sensitivity comparable to that of the best currently available fluorometric assays. The robustness and sensitivity of the method enable a wide range of applications, illustrated here by counting eukaryotic cells. Using widely available and inexpensive benchtop hardware, the approach provides a highly accessible low-tech microscale alternative to more expensive DNA detection and cell counting techniques.
机译:在旋转磁场中将DNA和超顺磁珠结合在一起,会产生视觉上引人注目的多部分聚集体,从而实现无标记的光学检测和DNA的定量检测(每微升范围在皮克级的水平)。可以通过简单分析放置在磁力搅拌器上的微流孔的光学图像,直接对生物样品中的DNA进行定量,而无需事先纯化DNA。聚集是由于离液剂的存在(聚集的非特异性触发因素)或与功能化珠上的寡核苷酸杂交(序列特异性)而驱动的DNA /珠相互作用。本文证明了定量DNA的灵敏度可与目前最好的荧光测定法媲美。该方法的鲁棒性和灵敏性可实现广泛的应用,此处通过对真核细胞计数来说明。使用广泛可用且价格便宜的台式硬件,该方法为更昂贵的DNA检测和细胞计数技术提供了高度可访问的低技术微型替代品。

著录项

  • 来源
    《Journal of the American Chemical Society》 |2012年第12期|p.5689-5696|共8页
  • 作者单位

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States,Center for Microsystems for the Life Sciences, University of Virginia, Charlottesville, Virginia 22904, United States,Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115;

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States,Center for Microsystems for the Life Sciences, University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States,Center for Microsystems for the Life Sciences, University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Mechanical Engineering, University of California, Santa Barbara, California 93106, United States;

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States,Department of Pathology,University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Electrical Engineering, University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Pathology,University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States,Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, Virginia 22904, United States ,Center for Microsystems for the Life Sciences, University of Virginia, Charlottesville, Virginia 22904, United States;

    Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States,Department of Mechanical and Aerospace Engineering, University of Virginia, Charlottesville, Virginia 22904, United States ,Department of Pathology,University of Virginia, Charlottesville, Virginia 22904, United States,Center for Microsystems for the Life Sciences, University of Virginia, Charlottesville, Virginia 22904, United States;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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