title: GSM1143101: [Genome-Seq] Old yeast genome replicate2 (MNase-Seq input); Saccharomyces cerevisiae; MNase-Seq
library_construction_protocol: The same number of purified young and old cells was subjected to genomic DNA isolation. The masterpureTM yeast DNA purification kit (cat. Nos. MPY80010 and MPY 80200) was used to isolate the genomic DNA.  Briefly, 300 ml of yeast cell lysis solution was added to each cell pellet. 1 ml of 5 mg/ml RNase A was added to each pellet. The cells were resuspended and incubated at 65°C for 15 minutes.  Samples were placed on ice for 5 minutes. 150 ml MPC protein precipitation reagent was added and vortexed. The cellular debris was pelleted by centrifugation for 10 min at 13,200 rpm. The supernatant was added to 500 ml isopropanol and the DNA was pelleted by centrifugation at 13,200 rpm for 10 minutes. The pellet was washed with 0.5 ml 70% ethanol. The ethanol was removed and the pellet was dried at room temperature. The isolated genomic DNA was fragmented using a Covaris sonicator to mimic the size of mononucleosomal DNA fragments for the size selection. The libraries were generated using a KAPA library preparation kit (KK8200, Illumina Series). During the library construction the same spike-in control described for MNase-seq was included and added proportional to cell number. DNA fragments were size selected between 200-400 bp after ligation of 121 bp adapters.
attributes.tag: GEO Accession
attributes.value: GSM1143101