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C-TALE, a new cost-effective method for targeted enrichment of Hi-C/3C-seq libraries

Author:
Golov, Arkadiy K., Ulianov, Sergey V., Luzhin, Artem V., Kalabusheva, Ekaterina P., Kantidze, Omar L., Flyamer, Ilya M., Razin, Sergey V., Gavrilov, Alexey A.
Source:
Methods 2019
ISSN:
1046-2023
Subject:
bacterial artificial chromosomes, chromatin, cost effectiveness, genes, genomics, high-throughput nucleotide sequencing, hybridization probes, vertebrates
Abstract:
Studies performed using Hi-C and other high-throughput whole-genome C-methods have demonstrated that 3D organization of eukaryotic genomes is functionally relevant. Unfortunately, ultra-deep sequencing of Hi-C libraries necessary to detect loop structures in large vertebrate genomes remains rather expensive. However, many studies are in fact aimed at determining the fine-scale 3D structure of comparatively small genomic regions up to several Mb in length. Such studies typically focus on the spatial structure of domains of coregulated genes, molecular mechanisms of loop formation, and interrogation of functional significance of GWAS-revealed polymorphisms. Therefore, a handful of molecular techniques based on Hi-C have been developed to address such issues. These techniques commonly rely on in-solution hybridization of Hi-C/3C-seq libraries with pools of biotinylated baits covering the region of interest, followed by deep sequencing of the enriched library. Here, we describe a new protocol of this kind, C-TALE (Chromatin TArget Ligation Enrichment). Preparation of hybridization probes from bacterial artificial chromosomes and an additional round of enrichment make C-TALE a cost-effective alternative to existing many-versus-all C-methods.
Agid:
6484596