GSM2800726: 4_1_CHIP_K9ac_1; Mus musculus; ChIP-Seq
Lysates were clarifed from the sonicated nuclei of day 5 EBs, and chromatin fragments were isolated with antibodies indicated. ChIP libraries were prepared using a Kapa Hyper Preparation kit (KAPA Biosystems, Wilmington, MA) protocol for Illumina Platforms. Briefly, for each library, 5ng of ChIP DNA was end-repaired and 3’-adenylated using a proprietary master mix, then ligated to the specific NexTflex adaptors from Bioo Scientific (Bioo Scientific, Austin, TX). The adaptor-ligated DNA was enriched using a KAPA Hyper Library Preparation kit (KAPA Biosystems, KK8502) with 5 cycles of PCR (1 cycle at 98°C for 45 seconds; 4 cycles of 98°C for 15 seconds, 60°C for 30 seconds, and 72°C for 30 seconds; 1 cycle at 72°C for 1 minute), then purified with AmpureXP beads (Beckman Coulter, A63881). The library quality was validated on a 2200 TapeStation from Agilent Technologies (Agilent, Santa Clara, CA). Concentrations of the libraries were determined using a Kapa Library Quantification Kit (KAPA Biosystems, KK4933) and loaded on cBOT (Illumina) at final concentration of 1.5nM for cluster generation, then sequenced with 50bp single-read on a HiSeq3000 sequencer (Illumina).