GSM3772980: Bud27-TAP Input 1; Saccharomyces cerevisiae; ChIP-Seq
Cell disruption was carried out by vigorously vortexing this sample in the presence of 200 µl of glass beads for 15 min. After disruption, lysis buffer was added to the cells extract to adjust 1.5 ml and then samples were subsequently sonicated by a Bioruptor sonicator (Diagenode) for 45 min at 30 s on/off intervals.   Immunoprecipitation was performed in duplicate using 200 µl of magnetic beads (Dynabeads Pan Mouse IgG, Invitrogen) and 300 µl of the sonicated sample. Washing and de-crosslinking followed the conventional chromatin immunoprecipitation procedure; supernatants were collected in the same tube and DNA was purified with the Nucleospin Extract II Purification kit (Macherey-Nagel). The eluted DNA was concentrated by lyophilization. Input and immunoprecipitated DNAs were sequenced in an Illumina HiSeq 2000 instrument Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit.