Cells were scraped and sonicated utilizing Diagenode Bioruptor at High setting and cycling 30 seconds ON, 30 seconds OFF. Ovol2-DNA complexes were isolated with lab generated anti-Ovol2 antibody specific against mouse. Libraries were prepared according to instructions for BiooScientific NEXTflex ChIP-Seq Library Systems. Specifically, DNA quality and fragment size was assessed by Qubit Sensitivity DNA assay, and the following library preparation steps were achieved with the following mixes: End repair of immunoprecipitated DNA with NEXTflex ChIP End Repair, 3' adenylation NEXTflex ChIP Adenylation mix, Adapter ligation with NEXTflex ChIP-Seq barcode kit containing barcodes to be used. PCR amplification used NEXTflex ChIP Primer Mix supplied in Bioo Scientific NEXTflex ChIP-Seq Barcode kit and amplified for the following program: 2min @ 98C, 30sec @ 98C, 30 sec @ 65C (repeated 16 cycles), 1min @ 72C, 4min @ 72C. Quality of libraries again assesses by Qubit DNA high Sensitivity.