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.
Keywords: on-chip CE; denaturant electrophoresis; DNA separation; peptide nucleic acid
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Kaneda, S.; Ono, K.; Fukuba, T.; Nojima, T.; Yamamoto, T.; Fujii, T. A Rapid Method for Optimizing Running Temperature of Electrophoresis through Repetitive On-Chip CE Operations. Int. J. Mol. Sci. 2011, 12, 4271-4281.
Kaneda S, Ono K, Fukuba T, Nojima T, Yamamoto T, Fujii T. A Rapid Method for Optimizing Running Temperature of Electrophoresis through Repetitive On-Chip CE Operations. International Journal of Molecular Sciences. 2011; 12(7):4271-4281.
Kaneda, Shohei; Ono, Koichi; Fukuba, Tatsuhiro; Nojima, Takahiko; Yamamoto, Takatoki; Fujii, Teruo. 2011. "A Rapid Method for Optimizing Running Temperature of Electrophoresis through Repetitive On-Chip CE Operations." Int. J. Mol. Sci. 12, no. 7: 4271-4281.