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Microfluidic Gene Arrays for Rapid Genomic Profiling

机译：用于快速基因组分析的微流体基因阵列

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Genomic analysis tools have recently become an indispensable tool for the evaluation of gene expression in a variety of experiment protocols. Two of the main drawbacks to this technology are the labor and time intensive process for sample preparation and the relatively long times required for target/probe hybridization. In order to overcome these two technological barriers we have developed a microfluidic chip to perform on chip sample purification and labeling, integrated with a high density genearray. Sample purification was performed using a porous polymer monolithic material functionalized with an oligo dT nucleotide sequence for the isolation of high purity mRNA. These purified mRNA's can then rapidly labeled using a covalent fluorescent molecule which forms a selective covalent bond at the N7 position of guanine residues. These labeled mRNA's can then released from the polymer monolith to allow for direct hybridization with oligonucletide probes deposited in microfluidic channel. To allow for rapid target/probe hybridization high density microarray were printed in microchannels. The channels can accommodate array densities as high as 4000 probes. When oligonucleotide deposition is complete, these channels are sealed using a polymer film which forms a pressure tight seal to allow sample reagent flow to the arrayed probes. This process will allow for real time target to probe hybridization monitoring using a top mounted CCD fiber bundle combination. Using this process we have been able to perform a multi-step sample preparation to labeled target/probe hybridization in less than 30 minutes. These results demonstrate the capability to perform rapid genomic screening on a high density microfluidic microarray of oligonucleotides.
机译：最近，基因组分析工具已成为评估各种实验方案中基因表达的必不可少的工具。该技术的两个主要缺点是样品制备的劳力和时间密集的过程以及靶标/探针杂交所需的相对较长的时间。为了克服这两个技术障碍，我们开发了一种微流控芯片，用于芯片上样品的纯化和标记，并集成了高密度基因阵列。使用经寡聚dT核苷酸序列功能化的多孔聚合物整体材料进行样品纯化，以分离高纯度mRNA。然后可以使用共价荧光分子快速标记这些纯化的mRNA，该共价荧光分子在鸟嘌呤残基的N7位形成选择性共价键。然后可以将这些标记的mRNA从聚合物整体中释放出来，以便与沉积在微流体通道中的寡核苷酸探针直接杂交。为了实现快速的靶标/探针杂交，在微通道中印制了高密度微阵列。这些通道可以容纳多达4000个探头的阵列密度。当寡核苷酸沉积完成后，使用聚合物膜密封这些通道，该膜形成压力密封，以允许样品试剂流到阵列的探针。该过程将允许实时目标使用顶部安装的CCD光纤束组合进行探针杂交监测。使用此过程，我们能够在不到30分钟的时间内完成多步骤的样品制备，以实现标记的靶标/探针杂交。这些结果证明了在寡核苷酸的高密度微流体微阵列上进行快速基因组筛选的能力。

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