Skip to Content
Merck
CN

Protocol for High-Throughput Analysis of Sister-Chromatids Contacts.

STAR protocols (2020-12-31)
Elena Espinosa, Yoshiharu Yamaichi, François-Xavier Barre
ABSTRACT

Sister chromatid interactions are a key step to ensure the successful segregation of sister chromatids after replication. Our knowledge about this phenomenon is mostly based on microscopy approaches, which have some constraints such as resolution limit and the impossibility of studying several genomic positions at the same time. Here, we present a protocol for Hi-SC2, a high-throughput sequencing-based method, to monitor sister chromatid contacts after replication at high resolution throughout the genome, which we applied to study cohesion in Vibrio cholerae. For complete details on the use and execution of this protocol, please refer to Espinosa et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
GenElute Bacterial Genomic DNA Kits, sufficient for 70 purifications
Sigma-Aldrich
L-(+)-Arabinose, ≥99% (GC)
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
Kanamycin sulfate from Streptomyces kanamyceticus, Animal Component-free
Sigma-Aldrich
Gentamicin sulfate salt, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Chloramphenicol, ≥98% (HPLC)