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  • iHi-C 2.0: A simple approach for mapping native spatial chromatin organisation from low cell numbers.

iHi-C 2.0: A simple approach for mapping native spatial chromatin organisation from low cell numbers.

Methods (San Diego, Calif.) (2019-07-10)
Athanasia Mizi, Eduardo Gade Gusmao, Argyris Papantonis
ABSTRACT

Genome organization is now understood to be tightly linked to all genomic functions. Thus, the high-resolution mapping of higher-order chromosomal structures via 3C-based approaches has become an integral tool for studying transcriptional and cell cycle regulation, signaling effects or disease onset. Nonetheless, 3C-based protocols are not without caveats, like dependencies on fixation conditions, restriction enzyme pervasiveness in crosslinked chromatin and ligation efficiency. To address some of these caveats, we describe here the streamlined iHi-C 2.0 protocol that allows for the genome-wide interrogation of native spatial chromatin contacts without a need for chemical fixation. This approach improves ligation efficiency and presents minimal material losses, and is thus suitable for analysing samples with limiting cell numbers. Following high throughput sequencing, iHi-C 2.0 generates high signal-to-noise and focal maps of the interactions within and between mammalian chromosomes under native conditions.

MATERIALS
Product Number
Brand
Product Description

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