2026-03-09T06:01:49Z https://hiroshima.repo.nii.ac.jp/oai

oai:hiroshima.repo.nii.ac.jp:02007289 2025-02-21T09:37:47Z 1730444907710

High-sensitive analysis of DNA fragments by capillary gel electrophoresis using transient isotachophoresis preconcentration and fluorescence detection Xu, Zhongqi Xu Zhongqi Esumi, Toshiaki Esumi Toshiaki Ikuta, Natsuki Ikuta Natsuki Hirokawa, Takeshi Hirokawa Takeshi open access Copyright (c) 2009 Elsevier B.V. Capillary gel electrophoresis DNA fragments Transient isotachophoresis Fluorescence detection Ethidium bromide 570 In this report aimed on further development of a high-sensitivity capillary gel electrophoresis (CCE) method for analysis of DNA fragments, we firstly explored online transient isotachophoresis (tITP) preconcentration combined with fluorescence detection (FD). The fluorescence signal (excitation: 488 nm; emission: 590 nm) was generated using the intercalating dye of ethidium bromide (EB). It was found when the leading electrolyte (LE) was injected behind the sample zone, such a special tITP mode has significant advantages to solve the bubble formation issue and to improve the analytical performance stability. Two standard DNA samples, a 50 bp DNA step ladder and the phi X174/HaeIII digest. were used to evaluate the qualitative and quantitative abilities of the tITP-FD approach. A highly diluted sample (10,000-fold in the water, e.g. the phi X174/HaeIII digest diluted from 500 mu g/ml to the 50 ng/ml level) was enriched and detected; the LOD was down to 0.09 ng/ml for the 72 bp fragment, apparently improved more than 1000-fold in comparison with UV detection. Although the RSD of peak areas (n = 3) was around 15.5% for the sample was electrokinetically injected, good linearity of peak area response showed that the proposed method is suitable for quantitative analysis. Elsevier Science BV 2009 eng journal article AO https://hiroshima.repo.nii.ac.jp/records/2007289 10.1016/j.chroma.2009.01.066 http://dx.doi.org/10.1016/j.chroma.2009.01.066 0021-9673 AA00695371 Journal of Chromatography A Journal of Chromatography A 1216 17 3602 3602 3605 https://hiroshima.repo.nii.ac.jp/record/2007289/files/JChromA_1216_3602.pdf 238.7 KB