Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

Is there any effective tool how to obtain short non-conding RNA (miRNA) in a sufficient amount and in a highest purity?

Autoři: Bílková Zuzana | Kročová Eliška | Smělá Denisa | Kupčík Rudolf | Macák Jan | Řehulka Pavel

Rok: 2018

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Is there any effective tool how to obtain short non-conding RNA (miRNA) in a sufficient amount and in a highest purity?	Current research in the field of molecular biology is focused mainly on regulation of gene expression. One of the great subjects of interest is short non-coding RNA. These molecules may act as transcriptional and post-transcriptional regulators of gene expression in a broad range of organisms. In the field of human molecular biology, short non-coding molecules called miRNAs in length 21 - 23 nucleotides are revealed frequently as acting in physiological but also in pathophysiological role. It is clear that the simple and robust method for isolation of miRNA from various biological materials is the most crucial step deeply supporting the research of miRNA. Currently, there are many more or less specialized methods for extraction of nucleic acids differing in the length but there is no efficient tool for separation and purification of short miRNA. In the frame of research revealing the role of miRNA in vivo in healthy/tumor cells, the proper separation method for profiling miRNA expression was needed. In addition to traditional phenol-chloroform extraction suitable for long polynucleotides, established but also new inorganic micro- or nanomaterials are tested and already applied for the purification and isolation of nucleic acids. Such materials are mostly based on SiO2 and they are suitable mainly for the isolation of long DNA/RNA molecules (Y.Bai et al., 2013; Shi R., et al., 2009). Several metal oxides (e.g. TiO2 or ZrO2) have been also proven as an effective capture material for selective enrichment of particular group of biopolymers, e.g. long DNA or phosphopeptides (A. Eriksson, et al., 2011). It is obvious that short RNA in length about 20 nucleotides exhibits similar structural and chemical characteristics as phosphopeptides. The goal of this work was to test the suitability of traditional and new TiO2-based nanomaterials for efficient and enough selective isolation of short non-coding miRNA molecules. Optimized protocol is included. Acknowledgements: This wor