Microfluidic gene arrays for rapid genomic profiling

Jay A.A. West; Kyle W. Hukari; Gary A. Hux; Timothy J. Shepodd

doi:10.1117/12.578619

8 December 2004 Microfluidic gene arrays for rapid genomic profiling

Jay A.A. West, Kyle W. Hukari, Gary A. Hux, Timothy J. Shepodd

Proceedings Volume 5591, Lab-on-a-Chip: Platforms, Devices, and Applications; (2004) https://doi.org/10.1117/12.578619
Event: Optics East, 2004, Philadelphia, Pennsylvania, United States

Abstract

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.

Citation Download Citation

Jay A.A. West, Kyle W. Hukari, Gary A. Hux, and Timothy J. Shepodd "Microfluidic gene arrays for rapid genomic profiling", Proc. SPIE 5591, Lab-on-a-Chip: Platforms, Devices, and Applications, (8 December 2004); https://doi.org/10.1117/12.578619

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