GSM1849301: MNase-seq in WT yeast grown in YPA, rep #1; Saccharomyces cerevisiae; MNase-Seq
MNase digestion assays were performed as described previously (Rando, O.J. (2010). Genome-wide mapping of nucleosomes in yeast. Methods in enzymology 470, 105-118.). Briefly, after collection, the cells were cross-linked with formaldehyde, the cell wall was digested with Zymolyase, and the chromatin was digested with a titration of MNase. The resulting DNA was electrophoresed on an agarose gel, and an ideal digestion in which most of the genome (>80%) was digested into a mono-nucleosome was chosen. Mononucleosomal DNA was isolated and end-repaired for library preparation. MNase-seq libraries were constructed with NEBNext® Ultra™ DNA Library Prep Kit for Illumina®. The DNA was size selected and ampliﬁed by PCR using the adaptor primers. The ampliﬁed DNA was then puriﬁed and used for cluster generation and sequencing on the Illumina® HiSeq or NextSeq system.
GEO
GSM1849301