A protocol to study three-dimensional genome structure in individual mutant preimplantation mouse embryos.
STAR Protoc
; 4(2): 102267, 2023 Apr 30.
Article
em En
| MEDLINE
| ID: mdl-37126445
ABSTRACT
Hi-C studies the three-dimensional structure of the genome by detecting genome-wide chromatin regions that are in spatial proximity within the nucleus. We developed single-blastocyst Hi-C in mutant mouse embryos to genotype them on sequence. We describe steps for embryo fixation and nuclei permeabilization, after which chromatin is digested and re-ligated having incorporated a biotin-labeled nucleotide at the ligation junction. After cross-link reversal, we then detail purification of immobilized chimeric DNA ligations, library generation, sequencing, and genome-wide analysis of interactions. For complete details on the use and execution of this protocol, please refer to Andreu et al. (2022).1.
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Ano de publicação:
2023
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Article