A parylene-based dual channel micro-electrophoresis system for rapid mutation detection via heteroduplex analysis

Sertan Sukas, Ayse Elif Erson, Cuneyt Sert, Haluk Kulah

Research output: Contribution to journalArticleAcademicpeer-review

14 Citations (Scopus)

Abstract

A new dual channel micro-electrophoresis system for rapid mutation detection based on heteroduplex analysis was designed and implemented. Mutation detection was successfully achieved in a total separation length of 250 μm in less than 3 min for a 590 bp DNA sample harboring a 3 bp mutation causing an amino acid change. Parylene-C was used as the structural material for fabricating the micro-channels as it provides conformal deposition, transparency, biocompatibility, and low background fluorescence without any surface treatment. A new dual channel architecture was derived from the traditional cross-channel layout by forming two identical channels with independent sample loading and waste reservoirs. The control of injected sample volume was accomplished by a new u-turn injection technique with pull-back method. The use of heteroduplex analysis as a mutation detection method on a cross-linked polyacrylamide medium provided accurate mutation detection in an extremely short length and time. The presence of two channels on the microchip offers the opportunity of comparing the sample to be tested with a desired control sample rapidly, which is very critical for the accuracy and reliability of the mutation analyses, especially for clinical and research purposes.

Original languageEnglish
Pages (from-to)3752-3758
Number of pages7
JournalElectrophoresis
Volume29
Issue number18
DOIs
Publication statusPublished - 2008
Externally publishedYes

Keywords

  • Heteroduplex analysis
  • Microchip electrophoresis
  • Mutation detection
  • Parylene-C

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