GSM7898868: IP_SNTB_HA_1; Aspergillus flavus; ChIP-Seq
For RNA-seq, total RNAs were extracted using TRIzol Reagent (Invitrogen, cat. NO 15596026) following the methods by Chomczynski et al. For ChiP-seq, the mycelium were cross-linked with 1% formaldehyde for 10 min at room temperature, after which 0.125 mol/L glycine was added, and the mixture was rested for 5 min to terminate the cross-linking reaction. Nuclei were collected by centrifuging the samples at 2,000 g for 5 min at 4 °C after being treated with lysis buffer (ECOTOP, ES-8523-500). To fragment chromatin DNA, nuclei were treated with nucleus lysis buffer and sonicated. For the lysis-sonicated chromatin, 10% was stored as “Input”, 90% was used in IP reactions with specific anti-HA magnetic beads (Beyotime, P2121, China). The Input DNA and IP DNA were extracted using the phenol-chloroform method. For ChiP-seq, high-throughput DNA sequencing libraries were prepared using a NEXTFLEX® ChIP Seq Kit (NOVA-514120, Bioo Scientific). The library products corresponding to 200-500 bp were enriched, quantified, and sequenced on a NovaSeq 6000 sequencer (Illumina, USA) using the PE150 model.