High-sensitive analysis of DNA fragments by capillary gel electrophoresis using transient isotachophoresis preconcentration and fluorescence detection

Journal of Chromatography A 1216 巻 17 号 3602-3605 頁 2009 発行
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タイトル ( eng )
High-sensitive analysis of DNA fragments by capillary gel electrophoresis using transient isotachophoresis preconcentration and fluorescence detection
作成者
Xu Zhongqi
Esumi Toshiaki
Ikuta Natsuki
収録物名
Journal of Chromatography A
1216
17
開始ページ 3602
終了ページ 3605
抄録
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.
著者キーワード
Capillary gel electrophoresis
DNA fragments
Transient isotachophoresis
Fluorescence detection
Ethidium bromide
NDC分類
化学工業 [ 570 ]
言語
英語
資源タイプ 学術雑誌論文
出版者
Elsevier Science BV
発行日 2009
権利情報
Copyright (c) 2009 Elsevier B.V.
出版タイプ Author’s Original(十分な品質であるとして、著者から正式な査読に提出される版)
アクセス権 オープンアクセス
収録物識別子
[ISSN] 0021-9673
[DOI] 10.1016/j.chroma.2009.01.066
[NCID] AA00695371
[DOI] http://dx.doi.org/10.1016/j.chroma.2009.01.066