Journal
ANALYTICAL CHEMISTRY
Volume 84, Issue 15, Pages 6366-6369Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac301586q
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Funding
- Defense Advanced Research Projects Agency [N660001-09-C-2082]
- Shustek Stanford Graduate Fellowship
- Directorate For Engineering
- Div Of Chem, Bioeng, Env, & Transp Sys [1159092] Funding Source: National Science Foundation
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We introduce an on-chip electrokinetic assay to perform high-sensitivity nucleic acid (NA) detection. This assay integrates electrokinetic sample focusing using isotachophoresis (ITP) with a background signal-removal strategy that employs photopatterened, DNA-functionalized hydrogels. In this multistage assay, ITP first enhances hybridization kinetics between target NAs and end-labeled complementary reporters. After enhanced hybridization, migration through a DNA-functionalized hydrogel region removes excess reporters through affinity interactions. We demonstrate our assay on microRNAs, an important class of low-abundance biomarkers. The assay exhibits 4 orders of magnitude dynamic range, near 1 pM detection limits starting from less than 100 fg of microRNA, and high selectivity for mature microRNA sequences, all within a 10 min run time. This new microfluiclic framework provides a unique quantitative assay for NA detection.
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