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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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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. |
format | Online Article Text |
id | pubmed-9407779 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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