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DNA-Enrichment Microfluidic Chip for Chromatin Immunoprecipitation

机译:用于染色质免疫沉淀的DNA富集微流控芯片

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摘要

Chromatin immunoprecipitation (ChIP) is a powerful and widely applied technique for detecting association of individual proteins with specific genomic regions; the technique requires several complex steps and is tedious. In this paper, we develop a microbead-packed microfluidic chip which eliminates most of the laborious, time-consuming, and skill-dependent processes of the ChIP procedure. A computational fluid dynamics model was established to analyze fluidic behavior in a microbead-packed microchannel. With the use of the new chip, a ChIP procedure was performed to purify the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) gene from human embryonic kidney cells (cell line 293). The ChIP capability of the microfluidic chip was evaluated and compared with that of a commercial assay kit; the precipitation performance of both methods was almost identical as shown by quantitative measurement of DNA. However, our chip offers the advantage of low resin volume, and the experimental time is greatly reduced. In addition, our method is less dependent on individual technical skill. Therefore, we expect that our microfluidic chip-based ChIP method will be widely used in DNA-, gene-, and protein-related research and will improve experimental efficiency.
机译:染色质免疫沉淀(ChIP)是一种功能强大且广泛应用的技术,可用于检测单个蛋白质与特定基因组区域的关联。该技术需要几个复杂的步骤并且繁琐。在本文中,我们开发了一种微珠包装的微流控芯片,该芯片消除了ChIP程序中大多数费力,耗时且依赖技能的过程。建立了计算流体动力学模型来分析微珠填充微通道中的流体行为。使用新芯片,进行了ChIP程序以从人胚胎肾细胞(细胞系293)中纯化GAPDH(3-磷酸甘油醛脱氢酶)基因。评估了微流控芯片的ChIP功能,并与商业化验试剂盒进行了比较。两种方法的沉淀性能几乎相同,如通过DNA定量测量所显示的。但是,我们的芯片具有树脂体积低的优点,并且大大减少了实验时间。此外,我们的方法较少依赖个人技术技能。因此,我们希望基于微流体芯片的ChIP方法将被广泛用于DNA,基因和蛋白质相关的研究中,并将提高实验效率。

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