GSM1846298: yAS38_ChIPSeq_untagged_veg_control; Saccharomyces cerevisiae; ChIP-Seq
The ChIP protocol used is described in Kuang, et al. 2014. Briefly, ~50 OD cells were fixed with 1% formaldehyde for 15 minutes at 25°C with shaking at 90 rpm, quenched with 0.125M glycine for 10 minutes at 25°C, washed with water, washed with 1X PBS, pelleted, and stored at –80°C. Briefly, the frozen pellet was resuspended in 0.45ml ChIP lysis buffer and lysed by bead beating with 12 cycles (1 minute on, 1 minute off). The sample was then sonicated for 20 cycles (30 seconds on, 30 seconds off) using a Bioruptor. For the ChIP of Sub1-3HA, the sample was incubated with 5µl of 0.4mg/ml anti-HA 12CA5 (Roche) overnight with rotation. The KAPA Hyper Prep Kit for Illumina platforms was used for constructing the sequencing library, following the manufacturer's protocol with 17 cycles for the amplification of the library. The samples for ChIP-Seq were pooled in equimolar amounts and sent for sequencing on the HiSeq 2500.