Progress achieved between 2014-2017 in the extraction and sample preparation of nucleic acid by isotachophoresis is reviewed in this paper. The isolation and purification of nucleic acids is very often compromised by a complex matrix such as blood and other bodily fluids, samples from the scene of crime, fossil samples, etc. While most of the common nucleic acids isolation techniques are based on extraction with inherent limitations with regard to quantitative results, isotachophoretic focusing is a quantitative process with a theoretically unlimited concentration factor. Since isotachophoresis belongs to less traditional approaches of nucleic acids purification, we present not only the latest developments in the application of isotachophoresis for the nucleic acids concentration but also a brief description of the principles of this method.
- MeSH
- automatizace MeSH
- DNA analýza MeSH
- elektrolyty MeSH
- hybridizace nukleových kyselin MeSH
- izotachoforéza metody trendy MeSH
- lidé MeSH
- mikro RNA analýza MeSH
- mikrofluidika MeSH
- nukleové kyseliny izolace a purifikace MeSH
- polymerázová řetězová reakce MeSH
- pufry MeSH
- tělesné tekutiny metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy 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.
High-temperature He loop is a new experimental device for investigation of structural materials for high-temperature gas-cooled reactors. In the first period of the test operation of the device organic compounds were detected in the loop. At low temperatures ethanol and acetone predominated, at higher temperatures more complex organic compounds were observed. The source of organic compounds is residual oil and degreasers from the production of the loop. After multiple flushing of the loop the concentration of organics were notably lower but not zero. On the basis of these findings a change in production of similar devices is recommended to prevent contamination of the device with organics.
1 grafický list ; 33 cm
Grafický list s portrétem pražského chirurga Ignáce Fritze (1778-1841).
- Konspekt
- Lékařské vědy. Lékařství
- NLK Obory
- lékařství
- chirurgie
- NLK Publikační typ
- grafické listy
- O autorovi
- Fritz, Ignác František, 1778-1841 Autorita
