共查询到20条相似文献,搜索用时 31 毫秒
1.
2.
Genome-scale ChIP-chip analysis using 10,000 human cells 总被引:2,自引:0,他引:2
3.
4.
5.
Chromatin immunoprecipitation (ChIP) is a powerful tool for the characterization of covalent histone modifications and DNA-histone interactions in vivo. The procedure includes DNA-histone cross-linking in chromatin, shearing DNA into smaller fragments, immunoprecipitation with antibodies against the histone modifications of interest, followed by PCR identification of associated DNA sequences. In this protocol, we describe a simplified and optimized version of ChIP assay by reducing the number of experimental steps and isolation solutions and shortening preparation times. We include a nuclear isolation step before chromatin shearing, which provides a good yield of high-quality DNA resulting in at least 15 mug of DNA from each immunoprecipitated sample (from 0.2 to 0.4 g of starting tissue material) sufficient to test > or =25 genes of interest. This simpler and cost-efficient protocol has been applied for histone-modification studies of various Arabidopsis thaliana tissues and is easy to adapt for other systems as well. 相似文献
6.
7.
8.
9.
The centromere is a multi-functional complex comprising centromeric DNA and a number of proteins. To isolate unidentified
centromeric DNA sequences, centromere-specific histone H3 variants (CENH3) and chromatin immunoprecipitation (ChIP) have been
utilized in some plant species. However, anti-CENH3 antibody for ChIP must be raised in each species because of its species
specificity. Production of the antibodies is time-consuming and costly, and it is not easy to produce ChIP-grade antibodies.
In this study, we applied a HaloTag7-based chromatin affinity purification system to isolate centromeric DNA sequences in
tobacco. This system required no specific antibody, and made it possible to apply a highly stringent wash to remove contaminated
DNA. As a result, we succeeded in isolating five tandem repetitive DNA sequences in addition to the centromeric retrotransposons
that were previously identified by ChIP. Three of the tandem repeats were centromere-specific sequences located on different
chromosomes. These results confirm the validity of the HaloTag7-based chromatin affinity purification system as an alternative
method to ChIP for isolating unknown centromeric DNA sequences. The discovery of more than two chromosome-specific centromeric
DNA sequences indicates the mosaic structure of tobacco centromeres. 相似文献
10.
11.
12.
Marking histone H3 variants: How,when and why? 总被引:2,自引:0,他引:2
13.
14.
15.
16.
17.
18.
Chaperoning the histone H3 family 总被引:1,自引:0,他引:1
19.