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Fabrication of two‐weir structure‐based packed columns for on‐chip solid‐phase extraction of DNA
Author(s) -
Zhong Runtao,
Liu Dayu,
Yu Linfen,
Ye Nannan,
Dai Zhongpeng,
Qin Jianhua,
Lin Bingcheng
Publication year - 2007
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.200600604
Subject(s) - microchannel , chromatography , etching (microfabrication) , materials science , elution , chip , solid phase extraction , extraction (chemistry) , packed bed , nanotechnology , layer (electronics) , chemistry , computer science , telecommunications
Abstract Microchip‐based packed column SPE of DNA was performed using the microfabricated two‐weir structure within a microchannel. We developed two methods to fabricate the two‐weir structured glass chips: a “two‐side etching/alignment” method and a simplified “one‐side/two‐step etching” method. The former method required a straightforward alignment step, while the latter approach comprised a simplified wet etching process using paraffin wax as the temporary protective layer. Both methods were convenient and rapid as compared to the previous approaches. Through a reversibly sealed bead‐introduction channel, beads can be fed into or out of the chip columns, thus enabling refreshment of the packing materials. Using the proposed chip columns, highly efficient λ‐DNA extractions (average recovery >80%) were performed with good chip‐to‐chip reproducibility (RSD <10%). The on‐chip SPE procedure was completed within 15 min at the flow rate of 3 μL/min and the bulk of the loaded DNA was eluted within a small volume of ∼8 μL. Application of the microchip‐based packed columns was demonstrated by purifying PCR‐amplifiable genomic DNA from human hepatocellular carcinoma (HepG2) cells and human whole blood samples.