DAP-seq of Flammulina filiformis
Firstly, F. filiformis genomic DNA library was built using a NEBNext DNA Library Prep Master Mix Set for Illumina (NEB, USA). Secondly, the Halo Tag expression plasmid of fftec1 was constructed by fusing the CDS of fftec1 into the pFN19K HaloTag T7 SP6 Flexi Vector (Promega, USA), then, the recombinant plasmid was transferred into TnT SP6 High-Yield Wheat Germ Protein Expression System (L3260, Promega) for fusion protein production. The Magne HaloTag Beads were used to purify the fusion protein, then the beads and protein mixture were directly incubated with 500 ng genomic DNA library with slow rotation for 1 h at room temperature. Finally, non-specific bound DNA fragments were removed by washing several times, and the purified DNA fragments were sequenced on Illumina NovaSeq 6000 with PE 150 method.