Document Detail


Optimization of encoded hydrogel particles for nucleic acid quantification.
MedLine Citation:
PMID:  19435332     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Daniel C Pregibon; Patrick S Doyle
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  81     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2009 Jun 
Date Detail:
Created Date:  2009-06-12     Completed Date:  2009-08-14     Revised Date:  2011-09-26    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4873-81     Citation Subset:  IM    
Affiliation:
Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
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MeSH Terms
Descriptor/Qualifier:
DNA / analysis*
Fluorescent Dyes / chemistry
Hydrogels / chemistry*
Microfluidic Analytical Techniques / instrumentation,  methods*
Nucleic Acid Hybridization
Polyethylene Glycols / chemistry
RNA / analysis*
Grant Support
ID/Acronym/Agency:
R21 EB008814-01A1/EB/NIBIB NIH HHS; R21EB008814/EB/NIBIB NIH HHS
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 0/Hydrogels; 0/Polyethylene Glycols; 63231-63-0/RNA; 9007-49-2/DNA
Comments/Corrections

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