title: GSM7467017: 2-2-MMS-12; Saccharomyces cerevisiae; OTHER
library_construction_protocol: 10 ml cell pellets were collected per conditions and were snap frozen at stored at -80°C. DNA was isolated from each cell pellet using Yeast Smash and Grab method. Briefly, to each cell pellet in an eppendorf tube 300 µL of glass beads, 200 µL of lysis buffer (10 mM Tris pH 8.0, 1 mM EDTA, 100 mM NaCl, 1% SDA, 2% Triton X-100) and 200 µL of 1:1 phenol chloroform was added.  The cells were vortexed for 2 mins followed by adding another 200 µL of TE pH 8.0 and continued vortexing for a few seconds.  The tubes were then centrifuged at 13,500 rpm for 5 mins after which the top aqueous layer was transferred to a fresh tube. Two volumes of 100% ethanol was added and the tubes were centrifuged at 13,500 rpm for 2-3 mins at RT to pellet the DNA.  The DNA pellet was washed with ice cold 70% ethanol.  The pellets were then dried and the DNA pellet was resuspended in 50 µL TE pH 8.0. The eluted DNA was then sonicated to using Covaris M220 sonicator to get an average fragment size of 300 bp.  The  sonicated DNA was end repaied followed by dA Tailing. The sonicated, endrepaired and dA Tailed DNA was subjected to library construction by attaching Solexa adaptors to the ends using T4 DNA ligase. 400 ng of the DNA was then subjectedto PCR amplification using KAPA Hotstart Ready Mix (KAPA) using a unique sequencing primer per sample.  The amplified DNA was then purified using AMPure beads to remove free adaptors.  The libraries were then sequenced on Illumina Hi-Seq 2500 platform  (150 bp paired-end reads).