Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine
We report procedures to allow incorporation and detection of 5-ethynyl-2'-deoxyuridine (EdU) in fission yeast, a thymidine analogue which has some technical advantages over use of bromode-oxyuridine. Low concentrations of EdU (1μM) are sufficient to allow detection of incorporation in cells exp...
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| Format: | Journal article |
| Language: | English |
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Oxford University Press
2011
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| _version_ | 1826268086317088768 |
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| author | Hua, H Kearsey, S |
| author_facet | Hua, H Kearsey, S |
| author_sort | Hua, H |
| collection | OXFORD |
| description | We report procedures to allow incorporation and detection of 5-ethynyl-2'-deoxyuridine (EdU) in fission yeast, a thymidine analogue which has some technical advantages over use of bromode-oxyuridine. Low concentrations of EdU (1μM) are sufficient to allow detection of incorporation in cells expressing thymidine kinase and human equilibrative nucleoside transporter 1 (hENT1). However EdU is toxic and activates the rad3-dependent checkpoint, resulting in cell cycle arrest, potentially limiting its applications for procedures which require labelling over more than one cell cycle. Limited DNA synthesis, when elongation is largely blocked by hydroxyurea, can be readily detected by EdU incorporation using fluorescence microscopy. Thus EdU should be useful for detecting early stages of S phase, or DNA synthesis associated with DNA repair and recombination. |
| first_indexed | 2024-03-06T21:04:12Z |
| format | Journal article |
| id | oxford-uuid:3be6e4cf-dbdb-4938-838a-147d280c4d59 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T21:04:12Z |
| publishDate | 2011 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | oxford-uuid:3be6e4cf-dbdb-4938-838a-147d280c4d592022-03-26T14:10:15ZMonitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridineJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:3be6e4cf-dbdb-4938-838a-147d280c4d59Zoological sciencesEnglishOxford University Research Archive - ValetOxford University Press2011Hua, HKearsey, SWe report procedures to allow incorporation and detection of 5-ethynyl-2'-deoxyuridine (EdU) in fission yeast, a thymidine analogue which has some technical advantages over use of bromode-oxyuridine. Low concentrations of EdU (1μM) are sufficient to allow detection of incorporation in cells expressing thymidine kinase and human equilibrative nucleoside transporter 1 (hENT1). However EdU is toxic and activates the rad3-dependent checkpoint, resulting in cell cycle arrest, potentially limiting its applications for procedures which require labelling over more than one cell cycle. Limited DNA synthesis, when elongation is largely blocked by hydroxyurea, can be readily detected by EdU incorporation using fluorescence microscopy. Thus EdU should be useful for detecting early stages of S phase, or DNA synthesis associated with DNA repair and recombination. |
| spellingShingle | Zoological sciences Hua, H Kearsey, S Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine |
| title | Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine |
| title_full | Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine |
| title_fullStr | Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine |
| title_full_unstemmed | Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine |
| title_short | Monitoring DNA replication in fission yeast by incorporation of 5-ethynyl-2'-deoxyuridine |
| title_sort | monitoring dna replication in fission yeast by incorporation of 5 ethynyl 2 deoxyuridine |
| topic | Zoological sciences |
| work_keys_str_mv | AT huah monitoringdnareplicationinfissionyeastbyincorporationof5ethynyl2deoxyuridine AT kearseys monitoringdnareplicationinfissionyeastbyincorporationof5ethynyl2deoxyuridine |