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Anal Chem. 1997 Sep 1;69(17):3451-7. doi: 10.1021/ac9703919.

Integrated capillary electrophoresis on flexible silicone microdevices:  analysis of DNA restriction fragments and detection of single DNA molecules on microchips.

Author information

1
Bioanalytical Research, Novartis Pharma AG, CH-4002 Basel, Switzerland.

Abstract

Microchips for integrated capillary electrophoresis systems were produced by molding a poly(dimethylsiloxane) (PDMS) silicone elastomer against a microfabricated master. The good adhesion of the PDMS devices on clean planar surfaces allows for a simple and inexpensive generation of networks of sealed microchannels, thus removing the constraints of elaborate bonding procedures. The performance of the devices is demonstrated with both fast separations of φX-174/HaeIII DNA restriction fragments labeled with the intercalating dye YOYO-1 and fluorescently labeled peptides. Detection limits in the order of a few zeptomoles (10(-)(21) mol) have been achieved for each injected DNA fragment, corresponding to a mass detection limit of ∼2 fg for the 603 base pair fragment. Single λ-DNA molecules intercalated with YOYO-1 at a base pair-to-dye ratio of 10:1 could be detected with an uncomplicated laser-induced fluorescence detection setup. High single-molecule detection efficiency (>50%) was achieved under electrophoretically controlled mass transport conditions in PDMS microchannels.

PMID:
21639267
DOI:
10.1021/ac9703919

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