GSM5113970: CEA17_polII_casp_1; Aspergillus fumigatus; ChIP-Seq
15mg of dry mycelia were lysed in 800 μl of FA lysis buffer for 3 min using a Bullet Blender (Next Advance), resuspended in 500 μl of FA lysis buffer and sonicated using the Qsonica Q800R at 100% amplitude with 10 seconds ON and 15 seconds OFF cycles for a total sonication time of 30 minutes. Approximately 2µg of the chromatin recovered immuno-precipitated using 2 ng of anti-RNA polymerase II (RNA-PolII) subunit antibody (clone 3E8, Millipore) or anti-HA (HA) antibody (F7, Santa Cruz Biotecnology) for 1.5 h, followed by incubation with ∼15 μl of packed Protein A Sepharose matrix (GE Healthcare) for another 1.5 h at room temperature. The Protein A beads were washed with increasing ionic strength solutions to remove the unspecific bindings. The DNA was eluted and de-crosslinked at 65 °C overnight. Library preparation was carried out using NEBNext® Ultra II Library Prep kit (Illumina, cat. no. E7645L) according to manufacturer's protocol.