Informace o publikaci
Preparative concentration of nucleic acids fragments by capillary isotachophoretic analyzer
| Autoři | |
|---|---|
| Rok publikování | 2018 |
| Druh | Článek v odborném periodiku |
| Časopis / Zdroj | Journal of Chromatography A |
| Fakulta / Pracoviště MU | |
| Citace | |
| Doi | http://dx.doi.org/10.1016/j.chroma.2018.03.021 |
| Klíčová slova | DNA; lsotachophoresis; Nucleic acids; Sample preparation |
| Popis | 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 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. (C) 2018 Elsevier B.V. All rights reserved. |
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