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Methods ; 58(3): 277-88, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22776363

ABSTRACT

Accumulating evidence demonstrates that the three-dimensional (3D) organization of chromosomes within the eukaryotic nucleus reflects and influences genomic activities, including transcription, DNA replication, recombination and DNA repair. In order to uncover structure-function relationships, it is necessary first to understand the principles underlying the folding and the 3D arrangement of chromosomes. Chromosome conformation capture (3C) provides a powerful tool for detecting interactions within and between chromosomes. A high throughput derivative of 3C, chromosome conformation capture on chip (4C), executes a genome-wide interrogation of interaction partners for a given locus. We recently developed a new method, a derivative of 3C and 4C, which, similar to Hi-C, is capable of comprehensively identifying long-range chromosome interactions throughout a genome in an unbiased fashion. Hence, our method can be applied to decipher the 3D architectures of genomes. Here, we provide a detailed protocol for this method.


Subject(s)
Chromosome Mapping/methods , Genome, Fungal , Saccharomyces cerevisiae/genetics , Animals , Biotinylation , Cross-Linking Reagents/chemistry , DNA Cleavage , DNA Restriction Enzymes/chemistry , DNA, Circular/chemistry , DNA, Circular/genetics , DNA, Circular/isolation & purification , DNA, Fungal/chemistry , DNA, Fungal/genetics , Formaldehyde/chemistry , Gene Library , Humans , Nucleic Acid Conformation , ROC Curve , Sequence Analysis, DNA
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