GSM1314112: MN_Scerevisiae_rpd3_S-phase; Saccharomyces cerevisiae; MNase-Seq
200 ml cultures at 1x107 cells/ml were collected in G1 and at 60 minutes in S-phase in the presence of 0.2 M HU for preparation of mononucleosomal DNA. Cells were treated with 10 mg of Zymolyase  20T during 10 minutes at 30 ºC to induce spheroplasts formation. Mononucleosomal fragments were generated by digesting DNA with 600 or 450 units/ml (WT and rpd3 strains respectively) of micrococcal nuclease (MNAse) at 37 ºC during 10 minutes. The amount of MNase was optimized experimentally for each strain to generate a 80:20 ratio of mononucleosomes to dinucleosomes, as described (Lantermann et al. 2010). Mononucleosomal DNA was recovered from 1.5% agarose gels and sequenced in an Illumina Genome Analyzer IIx using the single-end sequencing protocol. Mononucleosomal DNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM1314112