An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells
Summary: Leakage of mitochondrial or nuclear DNA into the cytosol can occur following viral infections, radiation damage, and some cancers. Here, we present an optimized protocol for isolating and quantifying cytosolic DNA from mammalian cells. We describe steps for collecting cytosolic fractions fr...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724000789 |
| _version_ | 1797299723635261440 |
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| author | Aminu S. Jahun Frederic Sorgeloos Ian G. Goodfellow |
| author_facet | Aminu S. Jahun Frederic Sorgeloos Ian G. Goodfellow |
| author_sort | Aminu S. Jahun |
| collection | DOAJ |
| description | Summary: Leakage of mitochondrial or nuclear DNA into the cytosol can occur following viral infections, radiation damage, and some cancers. Here, we present an optimized protocol for isolating and quantifying cytosolic DNA from mammalian cells. We describe steps for collecting cytosolic fractions from cells, extracting DNA using columns, and quantifying extracted DNA using qPCR. This straightforward protocol can be completed in as little as 5 hours, and allows for the identification of the source of DNA.For complete details on the use and execution of this protocol, please refer to Jahun et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-07T22:53:49Z |
| format | Article |
| id | doaj.art-ef8b9c16a73e4b018ae2bf3a8579047c |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-07T22:53:49Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-ef8b9c16a73e4b018ae2bf3a8579047c2024-02-23T05:00:48ZengElsevierSTAR Protocols2666-16672024-03-0151102913An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cellsAminu S. Jahun0Frederic Sorgeloos1Ian G. Goodfellow2Division of Virology, Department of Pathology, University of Cambridge, Addenbrooke’s Hospital Level 5, Hills Road, CB2 0QQ Cambridge, UK; Corresponding authorDivision of Virology, Department of Pathology, University of Cambridge, Addenbrooke’s Hospital Level 5, Hills Road, CB2 0QQ Cambridge, UK; Université catholique de Louvain, de Duve Institute, MIPA-VIRO 74-49, 74 Avenue Hippocrate, 1200 Brussels, BelgiumDivision of Virology, Department of Pathology, University of Cambridge, Addenbrooke’s Hospital Level 5, Hills Road, CB2 0QQ Cambridge, UK; Corresponding authorSummary: Leakage of mitochondrial or nuclear DNA into the cytosol can occur following viral infections, radiation damage, and some cancers. Here, we present an optimized protocol for isolating and quantifying cytosolic DNA from mammalian cells. We describe steps for collecting cytosolic fractions from cells, extracting DNA using columns, and quantifying extracted DNA using qPCR. This straightforward protocol can be completed in as little as 5 hours, and allows for the identification of the source of DNA.For complete details on the use and execution of this protocol, please refer to Jahun et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724000789CancerCell BiologyCell separation/fractionationCell-based AssaysImmunologyMolecular Biology |
| spellingShingle | Aminu S. Jahun Frederic Sorgeloos Ian G. Goodfellow An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells STAR Protocols Cancer Cell Biology Cell separation/fractionation Cell-based Assays Immunology Molecular Biology |
| title | An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells |
| title_full | An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells |
| title_fullStr | An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells |
| title_full_unstemmed | An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells |
| title_short | An optimized protocol for the extraction and quantification of cytosolic DNA in mammalian cells |
| title_sort | optimized protocol for the extraction and quantification of cytosolic dna in mammalian cells |
| topic | Cancer Cell Biology Cell separation/fractionation Cell-based Assays Immunology Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724000789 |
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