GSM8564793: R4_c93plus_INPUT; Homo sapiens; ChIP-Seq
U2OS/shUb/HA-Ub(WT) and U2OS/shUb/HA-Ub(K29R) cells and S2-DRSC cells were fixed with 1% formaldehyde for 10 min. Then, glycine was added to quench the reaction and cells were subsequently lysed. Sonication was performed on isolated nuclei using a Covaris E220 Evolution. For each ChIP reaction, 30 μg of U2OS chromatin was mixed with 0.5% sonicated Drosophila S2 chromatin and then immunoprecipitated with the corresponding antibody. Libraries were costructed using the KAPA Hyperprep kit and Illumina-compatible indexed adapters (IDT, 10005903) following manufacturer's instruction. For the amplification step, 6-7 PCR cycles were used. Subsequently, the DNA underwent size selection using Agencourt AMPure XP beads (Beckman Coulter, A63881) to obtain fragments between 200-700 bp. Libraries were validated using D5000 DNA ScreenTape on a TapeStation 4200.