Chapter title |
Analyzing the Dynamics of DNA Replication in Mammalian Cells Using DNA Combing
|
---|---|
Chapter number | 4 |
Book title |
DNA Replication
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2596-4_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2595-7, 978-1-4939-2596-4
|
Authors |
Marta Bialic, Vincent Coulon, Marjorie Drac, Thierry Gostan, Etienne Schwob, Bialic, Marta, Coulon, Vincent, Drac, Marjorie, Gostan, Thierry, Schwob, Etienne |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
France | 1 | 5% |
Unknown | 19 | 95% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 7 | 35% |
Researcher | 4 | 20% |
Student > Master | 3 | 15% |
Student > Bachelor | 1 | 5% |
Other | 1 | 5% |
Other | 2 | 10% |
Unknown | 2 | 10% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 7 | 35% |
Biochemistry, Genetics and Molecular Biology | 6 | 30% |
Arts and Humanities | 1 | 5% |
Computer Science | 1 | 5% |
Physics and Astronomy | 1 | 5% |
Other | 1 | 5% |
Unknown | 3 | 15% |