| Chapter title |
An in vitro DNA double-strand break repair assay based on end-joining of defined duplex oligonucleotides.
|
|---|---|
| Chapter number | 33 |
| Book title |
DNA Repair Protocols
|
| Published in |
Methods in molecular biology, January 2012
|
| DOI | 10.1007/978-1-61779-998-3_33 |
| Pubmed ID | |
| Book ISBNs |
978-1-61779-997-6, 978-1-61779-998-3
|
| Authors |
Kamal Datta, Shubhadeep Purkayastha, Ronald D. Neumann, Thomas A. Winters |
| Abstract |
DNA double-strand breaks (DSBs) are caused by endogenous cellular processes such as oxidative metabolism, or by exogenous events like exposure to ionizing radiation or other genotoxic agents. Repair of these DSBs is essential for the maintenance of cellular genomic integrity. In human cells, and cells of other higher eukaryotes, DSBs are primarily repaired by the nonhomologous end-joining (NHEJ) DSB repair pathway. Most in vitro assays that have been designed to measure NHEJ activity employ linear plasmid DNA as end-joining substrates, and such assays have made significant contributions to our understanding of the biochemical mechanisms of NHEJ. Here we describe an in vitro end-joining assay employing linear oligonucleotides that has distinct advantages over plasmid-based assays for the study of structure-function relationships between the proteins of the NHEJ pathway and synthetic DNA end-joining substrates possessing predetermined DSB configurations and chemistries. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
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|---|---|---|
| France | 1 | 5% |
| Unknown | 18 | 95% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 21% |
| Student > Bachelor | 2 | 11% |
| Researcher | 2 | 11% |
| Professor > Associate Professor | 2 | 11% |
| Student > Master | 1 | 5% |
| Other | 3 | 16% |
| Unknown | 5 | 26% |
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|---|---|---|
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| Social Sciences | 1 | 5% |
| Chemistry | 1 | 5% |
| Engineering | 1 | 5% |
| Other | 0 | 0% |
| Unknown | 4 | 21% |