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Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate

While microRNAs are considered as excellent biomarkers of various diseases, there are still several remaining challenges regarding their isolation. In this study, we aimed to design a novel RNA isolation method that would help to overcome those challenges. Therefore, we present a novel phenol/chloro...

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Detalles Bibliográficos
Autores principales: Smela, Denisa, Zelinkova, Eliska, Rehulka, Pavel, Bilkova, Zuzana, Kupcik, Rudolf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9407779/
https://www.ncbi.nlm.nih.gov/pubmed/36012112
http://dx.doi.org/10.3390/ijms23168848
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author Smela, Denisa
Zelinkova, Eliska
Rehulka, Pavel
Bilkova, Zuzana
Kupcik, Rudolf
author_facet Smela, Denisa
Zelinkova, Eliska
Rehulka, Pavel
Bilkova, Zuzana
Kupcik, Rudolf
author_sort Smela, Denisa
collection PubMed
description While microRNAs are considered as excellent biomarkers of various diseases, there are still several remaining challenges regarding their isolation. In this study, we aimed to design a novel RNA isolation method that would help to overcome those challenges. Therefore, we present a novel phenol/chloroform-free, low-cost method for miRNA extraction. Within this method, RNA is extracted from cell lysate with an isopropanol/water/NaCl system, followed by solid-phase extraction using TiO(2) microspheres to effectively separate short RNAs from long RNA molecules. We also demonstrated the pH-dependent selectivity of TiO(2) microspheres towards different sizes of RNA. We were able to regulate the size range of extracted RNAs with simple adjustments in binding conditions used during the solid-phase extraction.
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spelling pubmed-94077792022-08-26 Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate Smela, Denisa Zelinkova, Eliska Rehulka, Pavel Bilkova, Zuzana Kupcik, Rudolf Int J Mol Sci Communication While microRNAs are considered as excellent biomarkers of various diseases, there are still several remaining challenges regarding their isolation. In this study, we aimed to design a novel RNA isolation method that would help to overcome those challenges. Therefore, we present a novel phenol/chloroform-free, low-cost method for miRNA extraction. Within this method, RNA is extracted from cell lysate with an isopropanol/water/NaCl system, followed by solid-phase extraction using TiO(2) microspheres to effectively separate short RNAs from long RNA molecules. We also demonstrated the pH-dependent selectivity of TiO(2) microspheres towards different sizes of RNA. We were able to regulate the size range of extracted RNAs with simple adjustments in binding conditions used during the solid-phase extraction. MDPI 2022-08-09 /pmc/articles/PMC9407779/ /pubmed/36012112 http://dx.doi.org/10.3390/ijms23168848 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Smela, Denisa
Zelinkova, Eliska
Rehulka, Pavel
Bilkova, Zuzana
Kupcik, Rudolf
Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate
title Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate
title_full Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate
title_fullStr Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate
title_full_unstemmed Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate
title_short Phenol/Chloroform-Free TiO(2)-Based miRNA Extraction from Cell Lysate
title_sort phenol/chloroform-free tio(2)-based mirna extraction from cell lysate
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9407779/
https://www.ncbi.nlm.nih.gov/pubmed/36012112
http://dx.doi.org/10.3390/ijms23168848
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