Preparative concentration of nucleic acids fragments by capillary isotachophoretic analyzer
Language English Country Netherlands Media print-electronic
Document type Journal Article
PubMed
29559269
DOI
10.1016/j.chroma.2018.03.021
PII: S0021-9673(18)30314-5
Knihovny.cz E-resources
- Keywords
- DNA, Isotachophoresis, Nucleic acids, Sample preparation,
- MeSH
- DNA analysis MeSH
- Electricity MeSH
- Electrophoresis, Capillary instrumentation methods MeSH
- Isotachophoresis instrumentation methods MeSH
- Nucleic Acids analysis MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- DNA MeSH
- Nucleic Acids MeSH
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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