Concentrating Genomic Length DNA in a Microfabricated Array

Concentrating Genomic Length DNA in a Microfabricated Array
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DOI:
10.1103/physrevlett.114.198303
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发表时间:
2015-05-15
影响因子:
8.6
通讯作者:
Sturm, James C.
Sturm, James C.
中科院分区:
物理与天体物理1区
文献类型:
--
作者:
Chen, Yu;Abrams, Ezra S.;Sturm, James C.

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我们证明,一个微制造的凸点阵列可以集中基因组长度的DNA分子有效地在连续的,高流速,高达40 μ m/s,如果单分子DNA球具有足够大的剪切模量。剪切模量的增加通过使用聚乙二醇(PEG)衍生的耗尽力将DNA分子压缩至最小螺旋尺寸来实现。我们绘制出的甜蜜点,浓度发生,作为PEG浓度和流速的函数,使用理论分析和实验相结合。从酶促反应中纯化DNA用于下一代DNA测序文库将是这一发展的重要应用。
We demonstrate that a microfabricated bump array can concentrate genomic-length DNA molecules efficiently at continuous, high flow velocities, up to 40 mu m/s, if the single-molecule DNA globule has a sufficiently large shear modulus. Increase in the shear modulus is accomplished by compacting the DNA molecules to minimal coil size using polyethylene glycol (PEG) derived depletion forces. We map out the sweet spot, where concentration occurs, as a function of PEG concentration and flow speed using a combination of theoretical analysis and experiment. Purification of DNA from enzymatic reactions for next-generation DNA-sequencing libraries will be an important application of this development.