دورية أكاديمية
Analyzing the dynamics of DNA replication in Mammalian cells using DNA combing.
| العنوان: | Analyzing the dynamics of DNA replication in Mammalian cells using DNA combing. |
|---|---|
| المؤلفون: | Bialic M; Institute of Molecular Genetics, CNRS UMR5535 & Université Montpellier, 1919 route de Mende, Montpellier, 34293, France., Coulon V, Drac M, Gostan T, Schwob E |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2015; Vol. 1300, pp. 67-78. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | DNA Replication*, Biophysics/*methods , DNA/*metabolism , Mammals/*metabolism, Animals ; Embryo, Mammalian/cytology ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Image Processing, Computer-Assisted ; Mice ; Staining and Labeling |
| مستخلص: | How cells duplicate their chromosomes is a key determinant of cell identity and genome stability. DNA replication can initiate from more than 100,000 sites distributed along mammalian chromosomes, yet a given cell uses only a subset of these origins due to inefficient origin activation and regulation by developmental or environmental cues. An impractical consequence of cell-to-cell variations in origin firing is that population-based techniques do not accurately describe how chromosomes are replicated in single cells. DNA combing is a biophysical DNA fiber stretching method which permits visualization of ongoing DNA synthesis along Mb-sized single-DNA molecules purified from cells that were previously pulse-labeled with thymidine analogues. This allows quantitative measurements of several salient features of chromosome replication dynamics, such as fork velocity, fork asymmetry, inter-origin distances, and global instant fork density. In this chapter we describe how to obtain this information from asynchronous cultures of mammalian cells. |
| المشرفين على المادة: | 9007-49-2 (DNA) |
| تواريخ الأحداث: | Date Created: 20150429 Date Completed: 20160118 Latest Revision: 20150428 |
| رمز التحديث: | 20230127 |
| DOI: | 10.1007/978-1-4939-2596-4_4 |
| PMID: | 25916705 |
| قاعدة البيانات: | MEDLINE |
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