Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification
There is great demand for flexible biomolecule analysis platforms that can precisely quantify very low levels of multiple targets directly in complex biological samples. Herein we demonstrate multiplexed quantification of microRNAs (miRNAs) on encoded hydrogel microparticles with subfemtomolar sensi...
| 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/1721.1/79043 |
| _version_ | 1811073009141678080 |
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| author | Chapin, Stephen C. Doyle, Patrick S. |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Chapin, Stephen C. Doyle, Patrick S. |
| author_sort | Chapin, Stephen C. |
| collection | MIT |
| description | There is great demand for flexible biomolecule analysis platforms that can precisely quantify very low levels of multiple targets directly in complex biological samples. Herein we demonstrate multiplexed quantification of microRNAs (miRNAs) on encoded hydrogel microparticles with subfemtomolar sensitivity and single-molecule reporting resolution. Rolling circle amplification (RCA) of a universal adapter sequence that is ligated to all miRNA targets captured on gel-embedded probes provides the ability to label each target with multiple fluorescent reporters and eliminates the possibility of amplification bias. The high degree of sensitivity achieved by the RCA scheme and the resistance to fouling afforded by the use of gel particles are leveraged to directly detect miRNA in small quantities of unprocessed human serum samples without the need for RNA extraction or target-amplification steps. This versatility has powerful implications for the development of rapid, noninvasive diagnostic assays. |
| first_indexed | 2024-09-23T09:27:13Z |
| format | Article |
| id | mit-1721.1/79043 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:27:13Z |
| publishDate | 2013 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/790432022-09-30T14:26:46Z Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification Chapin, Stephen C. Doyle, Patrick S. Massachusetts Institute of Technology. Department of Chemical Engineering Chapin, Stephen C. Doyle, Patrick S. There is great demand for flexible biomolecule analysis platforms that can precisely quantify very low levels of multiple targets directly in complex biological samples. Herein we demonstrate multiplexed quantification of microRNAs (miRNAs) on encoded hydrogel microparticles with subfemtomolar sensitivity and single-molecule reporting resolution. Rolling circle amplification (RCA) of a universal adapter sequence that is ligated to all miRNA targets captured on gel-embedded probes provides the ability to label each target with multiple fluorescent reporters and eliminates the possibility of amplification bias. The high degree of sensitivity achieved by the RCA scheme and the resistance to fouling afforded by the use of gel particles are leveraged to directly detect miRNA in small quantities of unprocessed human serum samples without the need for RNA extraction or target-amplification steps. This versatility has powerful implications for the development of rapid, noninvasive diagnostic assays. National Institutes of Health (U.S.) (NIH Grant R21EB008814) Ragon Institute of MGH, MIT and Harvard 2013-05-30T18:04:54Z 2013-05-30T18:04:54Z 2011-08 2011-06 Article http://purl.org/eprint/type/JournalArticle 0003-2700 1520-6882 http://hdl.handle.net/1721.1/79043 Chapin, Stephen C., and Patrick S. Doyle. Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification. Analytical Chemistry 83, no. 18 (September 15, 2011): 7179-7185. en_US http://dx.doi.org/10.1021/ac201618k 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 | Chapin, Stephen C. Doyle, Patrick S. Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification |
| title | Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification |
| title_full | Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification |
| title_fullStr | Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification |
| title_full_unstemmed | Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification |
| title_short | Ultrasensitive Multiplexed MicroRNA Quantification on Encoded Gel Microparticles Using Rolling Circle Amplification |
| title_sort | ultrasensitive multiplexed microrna quantification on encoded gel microparticles using rolling circle amplification |
| url | http://hdl.handle.net/1721.1/79043 |
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