title: GSM1974378: RNR3 OE relicate 2 Input (Total); Saccharomyces cerevisiae; OTHER
library_construction_protocol: Genomic DNA was isolated with standard phenol chloroform extraction Genomic DNA isolation and shearing by sonication has been described previously (Knott et al., 2012) 1 µg total genomic DNA (sheared to ~300 bp average) was end-repaired and ligated with a barcoded, Illumina-compatible adapter using NEBNext Ultra End Repair/dA-Tailing Module (E7442) and NEBNext Ultra Ligation Module (E7445) (Dunham and Friesen, 2013).  AMPure beads (Beckman Coulter Cat#A63880) were used to isolate the genomic DNA and its concentration was determined by spectroscopy (Nanodrop).  Equal amounts of multiple such barcoded DNA samples were pooled; 20 ng was set aside as “Input” and 1 µg of this pool was subject to BrdU-IP as described previously (Knott et al., 2012).  The IP and Input DNA samples were PCR-amplified separately with indexed Illumina-compatible primers.  Amplified DNA was isolated using AMPure beads and validated and quantified on a Bioanalyzer, and pooled.
attributes.tag: GEO Accession
attributes.value: GSM1974378