Free flow isotachophoresis DNA purification in channel from disposable non-woven fabric bed

0509134 - ÚIACH 2020 PL eng A - Abstrakt
Duša, Filip - Moravcová, Dana - Šlais, Karel
Free flow isotachophoresis DNA purification in channel from disposable non-woven fabric bed.
CECE 2019. Gdańsk: University of Gdańsk, 2019. s. 44-44.
[CECE 2019. International Interdisciplinary Meeting on Bioanalysis /16./. 24.09.2019-26.09.2019, Gdańsk]
Grant CEP: GA ČR(CZ) GA19-00742S; GA MV VI20172020069
Institucionální podpora: RVO:68081715
Klíčová slova: DNA purification * free solution isotachophoresis * nucleic acids
Obor OECD: Analytical chemistry

Application of the discontinuous buffer systems for analysis of compounds in electrophoresis has been used in analytical methods for many decades. Currently, there has been a renewed interest in isotachophoresis due to its remarkable potential in extraction and focusing of nucleic acids. Our laboratory has been using free-flow electrophoresis in nonwoven beds for a number of electrophoretic modes of separation. In this work we propose free flow isotachophoresis in a strip of nonwoven fabric (approx. 4 x 170 x 1 mm) for purification and concentration of DNA. For analysis we used a lab-made platform published earlier [1]. The concentrating effect is presented using purified dsDNA extracted from salmon. Concentrating effect, determined with DNA gel electrophoresis, reached up to 50 times depending on the volume of a collected fraction. The part of the strip containing the DNA was marked by three dyes with mobilities selected to surround the mobility of dsDNA. The method was further tested for purification of dsDNA from yeast (Saccharomyces cerevisiae) cellular debris originating from sonication of yeast cells. The concentrating effect of the free flow isotachophoresis was reduced by broadening of dsDNA rich fraction caused by co-migrating simple organic acids coming from the sample matrix. Purification of dsDNA from proteins from sonicated yeast cells was observed with the proposed method.
Trvalý link: http://hdl.handle.net/11104/0299917