title: Chromosome Conformation Capture (3C) in Budding Yeast.
abstract: Chromosome conformation capture (3C) is a method for studying chromosomal organization that takes advantage of formaldehyde cross-linking to measure the spatial association of two pieces of chromatin. The 3C method begins with whole-cell formaldehyde fixation of chromatin. After cell lysis, solubilized chromatin is digested with a type II restriction endonuclease, and cross-linked DNA fragments are ligated together. Cross-links are reversed by degradation with proteinase K, and chimeric DNA molecules are purified by standard phenol:chloroform extraction. The resulting 3C library represents chromatin fragments that may be separated by large genomic distances or located on different chromosomes, but are close enough in three-dimensional space for cross-linking. Locus-specific oligonucleotide primers are used to detect interactions of interest in the 3C library using end-point polymerase chain reaction (PCR).
authors.affiliation: Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605.
authors.forename: Jon-Matthew
authors.initials: JM
authors.lastname: Belton
authors.affiliation: Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605.
authors.forename: Job
authors.initials: J
authors.lastname: Dekker
mesh_terms: D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D002875:Chromosomes / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000494:pharmacology; D005557:Formaldehyde / Q000494:pharmacology; D008968:Molecular Conformation; D004718:Saccharomycetales / Q000648:ultrastructure*
chemical_list: D002843:Chromatin; D003432:Cross-Linking Reagents; D005557:Formaldehyde