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- W2796386047 abstract "Formaldehyde cross-linking of DNA to associated proteins is a relatively straightforward method, but it is also the most critical step in the chromatin immunoprecipitation (ChIP) and 3C analyses. Although formaldehyde is a highly permeable cross-linker, its maximum cross-linking efficiencies are estimated to be at ∼1% for mammalian cells because reactivity is limited to amines. Therefore, a relatively large number of cells are required for 3C and ChIP-based assays. Five hundred million cross-linked diploid cells are equivalent to ∼1.66 fmol of the genome. Thus, only ∼100 amol of genomic copies is analyzed for one ChIP assay. Because the quality of cross-linked chromatin can vary, even when generated under near-identical conditions, it is preferable to generate multiple large batches. This protocol describes growing and cross-linking IMR90 primary human fibroblast cells for ChIP analysis. For other cell types, some modification of the protocol is necessary." @default.
- W2796386047 created "2018-04-13" @default.
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- W2796386047 date "2018-04-01" @default.
- W2796386047 modified "2023-10-14" @default.
- W2796386047 title "Formaldehyde Cross-Linking" @default.
- W2796386047 doi "https://doi.org/10.1101/pdb.prot082594" @default.
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