title: A rapid method for optimizing running temperature of electrophoresis through repetitive on-chip CE operations.
abstract: In this paper, a rapid and simple method to determine the optimal temperature conditions for denaturant electrophoresis using a temperature-controlled on-chip capillary electrophoresis (CE) device is presented. Since on-chip CE operations including sample loading, injection and separation are carried out just by switching the electric field, we can repeat consecutive run-to-run CE operations on a single on-chip CE device by programming the voltage sequences. By utilizing the high-speed separation and the repeatability of the on-chip CE, a series of electrophoretic operations with different running temperatures can be implemented. Using separations of reaction products of single-stranded DNA (ssDNA) with a peptide nucleic acid (PNA) oligomer, the effectiveness of the presented method to determine the optimal temperature conditions required to discriminate a single-base substitution (SBS) between two different ssDNAs is demonstrated. It is shown that a single run for one temperature condition can be executed within 4 min, and the optimal temperature to discriminate the SBS could be successfully found using the present method.
authors.affiliation: LIMMS-CNRS/IIS (UMI2820), Institute of Industrial Science, University of Tokyo, Tokyo 153-8505, Japan; E-Mail: shk@iis.u-tokyo.ac.jp.
authors.forename: Shohei
authors.initials: S
authors.lastname: Kaneda
authors.forename: Koichi
authors.initials: K
authors.lastname: Ono
authors.forename: Tatsuhiro
authors.initials: T
authors.lastname: Fukuba
authors.forename: Takahiko
authors.initials: T
authors.lastname: Nojima
authors.forename: Takatoki
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Teruo
authors.initials: T
authors.lastname: Fujii
mesh_terms: D004277:DNA, Single-Stranded / Q000737:chemistry / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D056656:Lab-On-A-Chip Devices; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature
chemical_list: D004277:DNA, Single-Stranded; D020135:Peptide Nucleic Acids