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Preparative concentration of nucleic acids fragments by capillary isotachophoretic analyzer

V. Datinská, I. Voráčová, J. Berka, F. Foret,

. 2018 ; 1548 (-) : 100-103. [pub] 20180315

Language English Country Netherlands

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc18024293

Sample preparation plays an important role in the DNA analysis workflow. Real samples often include a complex matrix, such as blood and other bodily fluids, or exogenous impurities, e.g., from the scene of crime. Most of the common nucleic acids isolation techniques are based on extractions; however, isotachophoretic focusing has recently attracted some interest for its simplicity and potential for very high enrichment factors and ease of automation. Here, we report on the use of a commercial isotachophoretic instrument for optimization of DNA focusing and preparative fraction collection. In order to achieve a high recovery and enrichment, experimental factors including electric current, sample amount and matrix were investigated experimentally as well as by computer simulation. The sample of a DNA ladder was injected in 30 μl volume and after ITP focusing the DNA zone was recovered using an on-column micropreparative collection valve. The DNA content in the collected sample was verified by fluorescence spectrometry and chip capillary electrophoresis with fluorescence detection.

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