Nucleic acid isolation and enrichment on a microchip

Jinho Kim, John P. Hilton, Kyung Ae Yang, Renjun Pei, Milan Stojanovic, Qiao Lin

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

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.

Original languageEnglish
Pages (from-to)183-190
Number of pages8
JournalSensors and Actuators, A: Physical
Volume195
DOIs
StatePublished - 1 Jun 2013

Keywords

  • Electrophoresis
  • Human immunoglobulin E
  • Microchannel
  • Solid-phase extraction
  • ssDNA

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