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DC Field | Value | Language |
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dc.contributor.author | Godwin, LS | - |
dc.contributor.author | Bridger, JM | - |
dc.contributor.author | Foster, HA | - |
dc.date.accessioned | 2020-11-30T14:48:55Z | - |
dc.date.available | 2020-11-30T14:48:55Z | - |
dc.date.issued | 2021-03-04 | - |
dc.identifier.citation | Godwin, L.S., Bridger, J.M. and Foster, H.A. (2021) 'Fluorescence In Situ Hybridization on DNA Halo Preparations to Reveal Whole Chromosomes, Telomeres and Gene Loci', Journal of Visualized Experiments (JoVE), (169), e62017, pp. 1 - 22 + streaming video.(09 minutes 27 seconds, available at: https://doi.org/10.3791/62017-v). doi: 10.3791/62017. | - |
dc.identifier.uri | https://bura.brunel.ac.uk/handle/2438/21931 | - |
dc.description | A streaming video (09 minutes 07 seconds) to accompany the text is available online at: https://www.jove.com/v/62017/fluorescence-situ-hybridization-on-dna-halo-preparations-to-reveal - doi: 10.3791/62017-v . | - |
dc.description.abstract | The genome is associated with several structures inside cell nuclei, in order to regulate its activity and anchor it in specific locations. These structures are collectively known as the nucleoskeleton and include the nuclear lamina, the nucleoli, and nuclear bodies. Although many variants of fluorescence in situ hybridization (FISH) exist to study the genome and its organization, these are often limited by resolution and provide insufficient information on the genome's association with nuclear structures. The DNA halo method uses high salt concentrations and nonionic detergents to generate DNA loops that remain anchored to structures within nuclei through attachment regions within the genome. Here, soluble nuclear proteins, such as histones, lipids, and DNA not tightly bound to the nuclear matrix, are extracted. This leads to the formation of a halo of unattached DNA surrounding a residual nucleus which itself contains DNA closely associated with internal nuclear structures and extraction-resistant proteins. These extended DNA strands enable increased resolution and can facilitate physical mapping. In combination with FISH, this method has the added advantage of studying genomic interactions with all the structures that the genome is anchored by. This technique, termed HALO-FISH, is highly versatile whereby DNA halos can be coupled with nucleic acid probes to reveal gene loci, whole chromosomes, alpha satellite, telomeres and even RNA. This technique provides an insight into nuclear organization and function in normal cells and in disease progression such as with cancer. | - |
dc.description.sponsorship | EU funded EURO-Laminopathies project and the Brunel Progeria Research Fund. | en_US |
dc.format.extent | 1 - 22 + streaming video (09 minutes 07 seconds available at: https://doi.org/10.3791/62017-v) | - |
dc.format.medium | Electronic | - |
dc.language.iso | en | en_US |
dc.publisher | MyJove Corporation | en_US |
dc.relation.uri | https://www.jove.com/video/62017 | - |
dc.relation.uri | https://www.jove.com/pdf-materials/62017 | - |
dc.rights | Copyright © The Author(s) 2021. Published by JoVE under a Creative Commons Attribution 3.0 License | - |
dc.rights.uri | https://creativecommons.org/licenses/by/3.0/ | - |
dc.subject | DNA halo preparation | en_US |
dc.subject | nuclear matrix | en_US |
dc.subject | nucleoskeleton | en_US |
dc.subject | genome organization | en_US |
dc.subject | DNA loops | en_US |
dc.title | Fluorescence in situ Hybridization on DNA Halo Preparations to reveal whole Chromosomes, Telomeres and Gene Loci | en_US |
dc.type | Article + Steaming Video | en_US |
dc.identifier.doi | https://doi.org/10.3791/62017 | - |
dc.relation.isPartOf | Journal of Vizualized Experiments | - |
pubs.publication-status | Published | - |
pubs.publisher-url | https://www.jove.com/video/62017 | - |
dc.identifier.eissn | 1940-087X | - |
dc.rights.holder | The Author(s) | - |
Appears in Collections: | Biological Sciences Dept of Life Sciences Research Papers |
Files in This Item:
File | Description | Size | Format | |
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FullText.pdf | Copyright © The Author(s) 2021. Published by JoVE under a Creative Commons Attribution 3.0 License | 726.84 kB | Adobe PDF | View/Open |
MaterialsSet.pdf | Copyright © The Author(s) 2021. Published by JoVE under a Creative Commons Attribution 3.0 License | 81.32 kB | Adobe PDF | View/Open |
This item is licensed under a Creative Commons License