Phenol/Chloroform-Free TiO<sub>2</sub>-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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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/16/8848 |
| _version_ | 1797409893990268928 |
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| author | Denisa Smela Eliska Zelinkova Pavel Rehulka Zuzana Bilkova Rudolf Kupcik |
| author_facet | Denisa Smela Eliska Zelinkova Pavel Rehulka Zuzana Bilkova Rudolf Kupcik |
| author_sort | Denisa Smela |
| collection | DOAJ |
| 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<sub>2</sub> microspheres to effectively separate short RNAs from long RNA molecules. We also demonstrated the pH-dependent selectivity of TiO<sub>2</sub> 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. |
| first_indexed | 2024-03-09T04:22:09Z |
| format | Article |
| id | doaj.art-9ba1e2df229c47e381d7ba40d930dd06 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T04:22:09Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9ba1e2df229c47e381d7ba40d930dd062023-12-03T13:46:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012316884810.3390/ijms23168848Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell LysateDenisa Smela0Eliska Zelinkova1Pavel Rehulka2Zuzana Bilkova3Rudolf Kupcik4Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech RepublicDepartment of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech RepublicDepartment of Molecular Pathology and Biology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech RepublicDepartment of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech RepublicDepartment of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, 532 10 Pardubice, Czech RepublicWhile 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<sub>2</sub> microspheres to effectively separate short RNAs from long RNA molecules. We also demonstrated the pH-dependent selectivity of TiO<sub>2</sub> 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.https://www.mdpi.com/1422-0067/23/16/8848miRNA isolationTiO<sub>2</sub>phenol/chloroform-freeshort RNA extraction |
| spellingShingle | Denisa Smela Eliska Zelinkova Pavel Rehulka Zuzana Bilkova Rudolf Kupcik Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell Lysate International Journal of Molecular Sciences miRNA isolation TiO<sub>2</sub> phenol/chloroform-free short RNA extraction |
| title | Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell Lysate |
| title_full | Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell Lysate |
| title_fullStr | Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell Lysate |
| title_full_unstemmed | Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell Lysate |
| title_short | Phenol/Chloroform-Free TiO<sub>2</sub>-Based miRNA Extraction from Cell Lysate |
| title_sort | phenol chloroform free tio sub 2 sub based mirna extraction from cell lysate |
| topic | miRNA isolation TiO<sub>2</sub> phenol/chloroform-free short RNA extraction |
| url | https://www.mdpi.com/1422-0067/23/16/8848 |
| work_keys_str_mv | AT denisasmela phenolchloroformfreetiosub2subbasedmirnaextractionfromcelllysate AT eliskazelinkova phenolchloroformfreetiosub2subbasedmirnaextractionfromcelllysate AT pavelrehulka phenolchloroformfreetiosub2subbasedmirnaextractionfromcelllysate AT zuzanabilkova phenolchloroformfreetiosub2subbasedmirnaextractionfromcelllysate AT rudolfkupcik phenolchloroformfreetiosub2subbasedmirnaextractionfromcelllysate |