Document Detail


Isotachophoresis with ionic spacer and two-stage separation for high sensitivity DNA hybridization assay.
MedLine Citation:
PMID:  23615527     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
We present an on-chip electrophoretic assay for rapid and high sensitivity nucleic acid (NA) detection. The assay uses isotachophoresis (ITP) to enhance NA hybridization and an ionic spacer molecule to subsequently separate reaction products. In the first stage, the probe and target focus and mix rapidly in free solution under ITP. The reaction mixture then enters a region containing a sieving matrix, which allows the spacer ion to overtake and separate the slower probe-target complex from free, unhybridized probes. This results in the formation of two focused ITP peaks corresponding to probe and probe-target complex signals. For a 149 nt DNA target, we achieve a 220 fM limit of detection (LOD) within 10 min, with a 3.5 decade dynamic range. This LOD constitutes a 12× improvement over previous ITP-based hybridization assays. The technique offers an alternative to traditional DNA hybridization assays, and can be multiplexed and extended to detect other biomolecules.
Authors:
Charbel Eid; Giancarlo Garcia-Schwarz; Juan G Santiago
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-4-25
Journal Detail:
Title:  The Analyst     Volume:  -     ISSN:  1364-5528     ISO Abbreviation:  Analyst     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-4-25     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372652     Medline TA:  Analyst     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Mechanical Engineering, Stanford University, 440 Escondido Mall, Bldg 530, room 225, Stanford, CA 94305, USA. juan.santiago@stanford.edu.
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