Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification
The accurate quantification of nucleic acids is of utmost importance for clinical diagnostics, drug discovery, and basic science research. These applications require the concurrent measurement of multiple targets while demanding high-throughput analysis, high sensitivity, specificity between closely...
| Main Authors: | , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Chemical Society
2013
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| Online Access: | http://hdl.handle.net/1721.1/79830 |
| _version_ | 1826193519567437824 |
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| author | Pregibon, Daniel C. Doyle, Patrick S. |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Pregibon, Daniel C. Doyle, Patrick S. |
| author_sort | Pregibon, Daniel C. |
| collection | MIT |
| description | The accurate quantification of nucleic acids is of utmost importance for clinical diagnostics, drug discovery, and basic science research. These applications require the concurrent measurement of multiple targets while demanding high-throughput analysis, high sensitivity, specificity between closely related targets, and a wide dynamic range. In attempt to create a technology that can simultaneously meet these demands, we recently developed a method of multiplexed analysis using encoded hydrogel particles. Here, we demonstrate tuning of hydrogel porosity with semi-interpenetrating networks of poly(ethylene glycol), develop a quantitative model to understand hybridization kinetics, and use the findings from these studies to enhance particle design for nucleic acid detection. With an optimized particle design and efficient fluorescent labeling scheme, we demonstrate subattomole sensitivity and single-nucleotide specificity for small RNA targets. |
| first_indexed | 2024-09-23T09:40:25Z |
| format | Article |
| id | mit-1721.1/79830 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:40:25Z |
| publishDate | 2013 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/798302022-09-30T16:05:08Z Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification Pregibon, Daniel C. Doyle, Patrick S. Massachusetts Institute of Technology. Department of Chemical Engineering Doyle, Patrick S. Pregibon, Daniel C. The accurate quantification of nucleic acids is of utmost importance for clinical diagnostics, drug discovery, and basic science research. These applications require the concurrent measurement of multiple targets while demanding high-throughput analysis, high sensitivity, specificity between closely related targets, and a wide dynamic range. In attempt to create a technology that can simultaneously meet these demands, we recently developed a method of multiplexed analysis using encoded hydrogel particles. Here, we demonstrate tuning of hydrogel porosity with semi-interpenetrating networks of poly(ethylene glycol), develop a quantitative model to understand hybridization kinetics, and use the findings from these studies to enhance particle design for nucleic acid detection. With an optimized particle design and efficient fluorescent labeling scheme, we demonstrate subattomole sensitivity and single-nucleotide specificity for small RNA targets. National Institute of Biomedical Imaging and Bioengineering (U.S.) (Grant R21EB008814) Massachusetts Institute of Technology (CSBi Merck Fellowship) Deshpande Center for Technological Innovation (Massachusetts Institute of Technology. School of Engineering) 2013-08-12T18:15:23Z 2013-08-12T18:15:23Z 2009-06 2009-03 Article http://purl.org/eprint/type/JournalArticle 0003-2700 1520-6882 http://hdl.handle.net/1721.1/79830 Pregibon, Daniel C., and Patrick S. Doyle. Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification. Analytical Chemistry 81, no. 12 (June 15, 2009): 4873-4881. en_US http://dx.doi.org/10.1021/ac9005292 Analytical Chemistry Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society PMC |
| spellingShingle | Pregibon, Daniel C. Doyle, Patrick S. Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification |
| title | Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification |
| title_full | Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification |
| title_fullStr | Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification |
| title_full_unstemmed | Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification |
| title_short | Optimization of Encoded Hydrogel Particles for Nucleic Acid Quantification |
| title_sort | optimization of encoded hydrogel particles for nucleic acid quantification |
| url | http://hdl.handle.net/1721.1/79830 |
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