title: Nucleic Acid Isolation and Enrichment on a Microchip.
abstract: This paper presents a microchip that isolates and enriches target-binding single-stranded DNA (ssDNA) from a randomized DNA mixture using a combination of solid-phase extraction and electrophoresis. Strands of ssDNA in a randomized mixture are captured via specific binding onto target-functionalized microbeads in a microchamber. The strands are further separated from impurities and enriched on-chip via electrophoresis. The microchip consists of two microchambers that are connected by a channel filled with agarose gel. In the isolation chamber, beads functionalized with human immunoglobulin E (IgE) are retained by a weir structure. An integrated heater elevates the temperature in the chamber to elute desired ssDNA from the beads, and electrophoretic transport of the DNA through the gel to the second chamber is accomplished by applying an electric potential difference between the two chambers. Experimental results show that ssDNA expressing binding affinity to IgE was captured and enriched from a sample of ssDNA with random sequences, demonstrating the potential of the microchip to enhance the sensitivity of ssDNA detection methods in dilute and complex biological samples.
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY 10027.
authors.forename: Jinho
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY 10027.
authors.forename: John P
authors.initials: JP
authors.lastname: Hilton
authors.affiliation: Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, New York, NY 10032.
authors.forename: Kyung A
authors.initials: KA
authors.lastname: Yang
authors.affiliation: Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, New York, NY 10032.
authors.forename: Renjun
authors.initials: R
authors.lastname: Pei
authors.affiliation: Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, New York, NY 10032.
authors.forename: Milan
authors.initials: M
authors.lastname: Stojanovic
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY 10027.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Lin
keywords: Solid-phase extraction; electrophoresis; human immunoglobulin E; microchannel; ssDNA
references.citation: J Chromatogr A. 2002 Jul 19;963(1-2):3-17
references.pmid: 12187984
references.citation: Talanta. 2007 Dec 15;74(3):342-57
references.pmid: 18371648
references.citation: Electrophoresis. 1999 Jul;20(10):1962-77
references.pmid: 10451104
references.citation: J Immunol. 1996 Jul 1;157(1):221-30
references.pmid: 8683119
references.citation: Methods Mol Med. 2007;140:291-307
references.pmid: 18085215
references.citation: Anal Chem. 2007 Mar 1;79(5):2168-73
references.pmid: 17269791
references.citation: Chem Soc Rev. 2010 Mar;39(3):912-22
references.pmid: 20179814
references.citation: Talanta. 2002 Feb 11;56(2):233-66
references.pmid: 18968499
references.citation: Anal Chem. 2007 Aug 15;79(16):6201-7
references.pmid: 17616169
references.citation: J Am Chem Soc. 2003 Oct 29;125(43):13026-7
references.pmid: 14570466
references.citation: Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1229-33
references.pmid: 1741376
references.citation: J Chromatogr A. 2000 Dec 1;902(1):91-105
references.pmid: 11192163
references.citation: Comput Biol Chem. 2007 Feb;31(1):11-35
references.pmid: 17218151
references.citation: Anal Chem. 2000 Feb 1;72(3):585-90
references.pmid: 10695146
references.citation: Anal Chem. 2007 Nov 1;79(21):8316-22
references.pmid: 17883279
references.citation: Anal Chim Acta. 2009 Oct 12;652(1-2):22-31
references.pmid: 19786168
references.citation: Anal Chim Acta. 2005 Aug 1;546(1):22-29
references.pmid: 29569551
references.citation: Nano Lett. 2010 Mar 10;10(3):765-72
references.pmid: 20151696
references.citation: Electrophoresis. 2000 May;21(8):1464-70
references.pmid: 10832874
references.citation: Anal Chem. 2007 May 15;79(10):3724-33
references.pmid: 17428034
references.citation: Anal Chem. 2006 Jun 15;78(12):3887-908
references.pmid: 16771530
references.citation: Anal Chem. 2009 Apr 15;81(8):2832-9
references.pmid: 19298056
references.citation: J Chromatogr A. 2008 Mar 14;1184(1-2):542-59
references.pmid: 18207148
references.citation: J Chromatogr A. 2005 Feb 25;1066(1-2):197-203
references.pmid: 15794571
references.citation: Anal Chem. 2006 Jul 15;78(14):4779-85
references.pmid: 16841895
references.citation: Science. 1986 Dec 19;234(4783):1582-5
references.pmid: 3538420
references.citation: Electrophoresis. 2007 Aug;28(16):2920-6
references.pmid: 17640088
references.citation: Sens Actuators B Chem. 2011 Jul 5;155(1):58-66
references.pmid: 21765612
references.citation: J Sep Sci. 2009 Aug;32(15-16):2752-8
references.pmid: 19585530
references.citation: Lab Chip. 2012 Jan 7;12(1):99-108
references.pmid: 22045330