alias: GSM3304632
title: GSM3304632: fun30 depletion, 0h; Saccharomyces cerevisiae; MNase-Seq
library_construction_protocol: Cells were fixed with 1% formaldehyde for 15 minutes. The reaction was stopped by adding glycine to a final concentration of 125 mM, and cells were harvested by centrifugation and washed. Pellet were kept in -80oC. Yeast cells were resuspended in Zymolyase buffer and treated with zymolyase (6U/od) and incubated for 25 min at 30 °C to generate spheroplasts. cells were centrifuged, washed and resuspended NP buffer. Lysed Spheroplasts were treated with MNAse (Micrococcal nuclease, worthington). MNAse concentration was calibrated to generate 80% mono nucleosomes (1 unit for 2.5od, 37C, 20m). The reactions were stopped by adding 100 μl of 5% SDS and 50 mM EDTA.   MNase digested chromatin was reverse cross linked, and MNase sequencing libraries were prepared (Blecher-Gonen et al. 2013).
identifiers.namespace: GEO
identifiers.id: GSM3304632