Abstract
Digestion of chromatin by micrococcal nuclease MNase followed by high throughput sequencing allows us to determine the location and occupancy of nucleosomes on the genome. Here in this protocol we have described optimized conditions of MNase digestion of filamentous fungus Neurospora crassa chromatin without a requirement of a nuclear fractionation step.
Keywords: Neurospora crassa, Nucleosome occupancy, MNase digestion
Materials and Reagents
Equipment
Procedure
Representative data
Figure 1. Representative image of MNase digested and purified Neurospora nucleosomal DNA loaded on 1.7% agarose gel
Recipes
Acknowledgments
This protocol was used for the work previously published in PLoS Genetics (Sancar et al., 2015). This work was supported by grants of the Deutsche Forschungsgemeinschaft: GRK1188 and SFB1036.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.