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On‐chip coupling of free‐solution transient ITP and CGE for highly efficient separation of dsDNA with variable sample loading amounts
Author(s) -
Liu Dayu,
Chen Bin,
Wang Lihui,
Zhou Xiaomian
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900333
Subject(s) - isotachophoresis , chemistry , electrolyte , computable general equilibrium , analytical chemistry (journal) , detection limit , chromatography , fluorescence , ion , coupling (piping) , materials science , physics , electrode , quantum mechanics , economics , metallurgy , macroeconomics , organic chemistry
We developed an on‐chip DNA analysis method, in which free‐solution transient isotachophoresis (FstITP) were coupled with CGE. Using chloride ions in the sample matrix and HEPES in the background electrolyte, respectively, as the leading and terminating ions, dsDNAs were isotachophoretically preconcentrated in free‐solution and then separated in sieving polymer. The coupling of FstITP and CGE enabled higher separation efficiency due to higher preconcentration rate in free‐solution. The FstITP‐CGE analysis offered adjustable signal intensities by varying sample injection time. With the maximum sample loading volume, the LOD of the FstITP‐CGE analysis was determined to be 0.24 ng/mL by confocal laser‐induced fluorescence detection. The FstITP‐CGE method is simple, robust and flexible, thus well suited to the analysis of highly saline DNA samples at different concentration level.