ZmO2 DAP-Seq of 14DAP endosperm from second biological zmros1ab mutant (control)
DNA from WT and zmros1ab mutant endosperm was was extracted using CTAB method. ZmO2 protein was expressed using TNT SP6 Wheat Germ Master Mix following manufacturers instruction. A total of 10 g genomic DNA was fragmented to ~200 bp, and adapters were ligated using NEXTflex Rapid DNA-Seq Kit. Next, the DNA was incubated with ZmO2 protein, and DNA bound by ZmO2 was PCR amplified using KAPA HiFi HotStart ReadyMix PCR Kit. The purified libraries were sequenced on Illumina NovaSeq 6000 under the paired-end mode.