Use of the combing process for the identification of DNA origins of replication
US6248537A · kind A · utility
Assignees
Inventors
Key dates
| Filing date | May 26, 2000 |
| Grant date | Jun 19, 2001 |
| Priority date | — |
| Expiry date | May 26, 2020 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6804
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Eukaryotic genomes are duplicated by the activation of multiple bidirectional origins of replication. The replication programs of these cells depend on the temporal and spatial organization of replication origins throughout the genome. To investigate the replication program in a higher eukaryote, we employed a technique called molecular combing. This technique allows for a quantitative analysis of DNA replication on a genome wide basis. As a model system, Xenopus laevis sperm chromatin were differentially labeled at successive time points after the beginning of DNA synthesis. Genomic DNA was then extracted and combed on a glass surface. Direct measurements made on the labeled DNA provided a comprehensive analysis of the spatial and temporal organization of the X. laevis early embryo replication program and revealed that the number of replication origins activated per kilobase increases throughout the period of DNA synthesis. local freq is 1/14 kb global is an increase of 5 fold per 100 kb each time point
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.