Fabrication of Uniform DNA-Conjugated Hydrogel Microparticles via Replica Molding for Facile Nucleic Acid Hybridization Assays

We identify and investigate several critical parameters in the fabrication of single-stranded DNA conjugated poly(ethylene glycol) (PEG) microparticles based on replica molding (RM) for highly uniform and robust nucleic acid hybridization assays. The effects of PEG-diacrylate, probe DNA, and photoin...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2010-07, Vol.82 (13), p.5851-5858
Main Authors: Lewis, Christina L, Choi, Chang-Hyung, Lin, Yan, Lee, Chang-Soo, Yi, Hyunmin
Format: Article
Language:English
Subjects:
Acrylates - chemistry
Analytical chemistry
Chemistry
Deoxyribonucleic acid
DNA
DNA - chemistry
DNA Probes - chemistry
Exact sciences and technology
Fluorescence
Fluorescent Dyes - chemistry
Food Microbiology
Hydrogels - chemistry
Immunoassay
Microscopy, Fluorescence
Oligonucleotide Array Sequence Analysis - methods
Polyethylene glycol
Polyethylene Glycols - chemistry
Spectrometric and optical methods
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Summary:We identify and investigate several critical parameters in the fabrication of single-stranded DNA conjugated poly(ethylene glycol) (PEG) microparticles based on replica molding (RM) for highly uniform and robust nucleic acid hybridization assays. The effects of PEG-diacrylate, probe DNA, and photoinitiator concentrations on the overall fluorescence and target DNA penetration depth upon hybridization are examined. Fluorescence and confocal microscopy results illustrate high conjugation capacity of the probe and target DNA, femtomole sensitivity, and sequence specificity. Combined, these findings demonstrate a significant step toward simple, robust, and scalable procedures to manufacture highly uniform and high-capacity hybridization assay particles in a well-controlled manner by exploiting many advantages that the batch processing-based RM technique offers. We envision that the results presented here may be readily applied to rapid and high-throughput hybridization assays for a wide variety of applications in bioprocess monitoring, food safety, and biological threat detection.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac101032r