title: GSM8163416: W22_ZmNAC128_ChIP1; Zea mays; ChIP-Seq
library_construction_protocol: About 2 g of kernel powder was resuspended in pre-cold  M1 buffer (10 mM sodium phosphate[pH 7], 0.1 M NaCl, 1 M 2-methyl 2,4-pentanediol, 10 mM β-mercaptoethanol and complete protease inhibitor cocktail). Cell nuclei were extracted using pre-cold M2 buffer (like M1 buffer with 10 mM MgCl2 and 0.5% Triton X-100),  and M3 buffer (like M1 buffer without 2-methyl 2,4-pentanediol), then washed 5 times separately. Cell nuclei were incubated with ZmNAC128, ZmNAC130 and IgG antibodies (final concentration, 20 μg/mL) for 12 h at 4℃. Then supernatant were incubated with dynabeads Protein A (Invitrogen) for 2 h at 4℃. After washing the samples four times, they were incubated with Tn5 transposome and tagmentation buffer (10 mM TAPS-NaOH (pH 8.5), 10 mM MgCl2, 10% (v/v) DMF, 0.5 mM spermidine, 0.05% (v/v) digitonin, and 1% (v/v) plant cocktail) at 37℃ for 5 min.  The reactions were stopped and then the protein-DNA complexes were eluted from the beads. Finally, after proteinase k (Invitrogen) treatment, ChIP-DNA libraries were constructed and sequenced using an Illumina NovaSeq 6000 platform (Illumina, San Diego, CA, USA) by BerryGenomics (China).