To interrogate the full spectrum of chromatin folding ranging from the level of nucleosomes to full chromosomes in mammals, we developed an enhanced 3C-based method called Micro-C. The protocol employs Micrococcal nuclease (MNase) to fragment the genome, which overcomes the resolution limit of ...
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a Schematic of Micro-C protocol. Nuclei were extracted from plant tissue and then separately cross-linked by formaldehyde (FA) and ethylene glycol bis (succinimidyl succinate) (EGS). Subsequently, chromatin was digested with micrococcal nuclease (MNase), and DNA overhangs were filled with biotin-dN...
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(a) Representative MNase titration DNA gel indicating the ideal level of digestion by MNase, based on the ratio of fragment sizes, for the RCMC protocol. (b) Representative size-selection gel for the RCMC protocol showing the dinucleosomal band that is extracted to obtain ligated fragments. (c...
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