Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives
Abstract The comet assay (CA), originally developed as toxicity test, quantifies DNA integrity from DNA distribution across an electric field. Compromised DNA moves across electric fields faster than intact DNA strands, leaving quantifiable signatures that resemble comet tails. Comet tail dimensions...
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| Format: | Article |
| Language: | English |
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Wiley
2025-02-01
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| Series: | Methods in Ecology and Evolution |
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| Online Access: | https://doi.org/10.1111/2041-210X.14493 |
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| author | Rejin Salimraj Alessio Perotti Marcin W. Wojewodzic Dagmar Frisch |
| author_facet | Rejin Salimraj Alessio Perotti Marcin W. Wojewodzic Dagmar Frisch |
| author_sort | Rejin Salimraj |
| collection | DOAJ |
| description | Abstract The comet assay (CA), originally developed as toxicity test, quantifies DNA integrity from DNA distribution across an electric field. Compromised DNA moves across electric fields faster than intact DNA strands, leaving quantifiable signatures that resemble comet tails. Comet tail dimensions reflect relative DNA damage. We optimised the CA protocol for individual dormant propagules (single‐egg comet assay or SE‐CA) to inform downstream analyses such as DNA sequencing of the DNA quality contained in natural genetic archives of past populations. As a model, we used dormant eggs of the microcrustacean Daphnia. We evaluated the feasibility of the SE‐CA protocol to quantify DNA damage caused by processing and storage conditions for dormant eggs and in dormant eggs retrieved from recently deposited to centuries‐old lake sediment. In conclusion, our protocol provides a cost‐effective method of assessing DNA damage in sedimentary propagules such as dormant Daphnia eggs. More generally, the SE‐CA is applicable to testing DNA integrity in individual propagules prior to genome sequencing or to quantify environmental impacts on natural sedimentary biobanks. |
| format | Article |
| id | doaj-art-4e587d1652dd4fcca4f34337543abf02 |
| institution | Kabale University |
| issn | 2041-210X |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Methods in Ecology and Evolution |
| spelling | doaj-art-4e587d1652dd4fcca4f34337543abf022025-02-05T05:43:20ZengWileyMethods in Ecology and Evolution2041-210X2025-02-0116229530110.1111/2041-210X.14493Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchivesRejin Salimraj0Alessio Perotti1Marcin W. Wojewodzic2Dagmar Frisch3Pharmacovigilance United BioSource Corporation Geneva SwitzerlandSchool of Biosciences The University of Birmingham Birmingham UKSchool of Biosciences The University of Birmingham Birmingham UKSchool of Biosciences The University of Birmingham Birmingham UKAbstract The comet assay (CA), originally developed as toxicity test, quantifies DNA integrity from DNA distribution across an electric field. Compromised DNA moves across electric fields faster than intact DNA strands, leaving quantifiable signatures that resemble comet tails. Comet tail dimensions reflect relative DNA damage. We optimised the CA protocol for individual dormant propagules (single‐egg comet assay or SE‐CA) to inform downstream analyses such as DNA sequencing of the DNA quality contained in natural genetic archives of past populations. As a model, we used dormant eggs of the microcrustacean Daphnia. We evaluated the feasibility of the SE‐CA protocol to quantify DNA damage caused by processing and storage conditions for dormant eggs and in dormant eggs retrieved from recently deposited to centuries‐old lake sediment. In conclusion, our protocol provides a cost‐effective method of assessing DNA damage in sedimentary propagules such as dormant Daphnia eggs. More generally, the SE‐CA is applicable to testing DNA integrity in individual propagules prior to genome sequencing or to quantify environmental impacts on natural sedimentary biobanks.https://doi.org/10.1111/2041-210X.14493bioarchiveDaphniaDNA degradationpaleogenomicsresurrection ecologysedimentary propagules |
| spellingShingle | Rejin Salimraj Alessio Perotti Marcin W. Wojewodzic Dagmar Frisch Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives Methods in Ecology and Evolution bioarchive Daphnia DNA degradation paleogenomics resurrection ecology sedimentary propagules |
| title | Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives |
| title_full | Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives |
| title_fullStr | Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives |
| title_full_unstemmed | Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives |
| title_short | Single‐egg comet assay: A protocol to quantify DNA damage in natural bioarchives |
| title_sort | single egg comet assay a protocol to quantify dna damage in natural bioarchives |
| topic | bioarchive Daphnia DNA degradation paleogenomics resurrection ecology sedimentary propagules |
| url | https://doi.org/10.1111/2041-210X.14493 |
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