GSM6774491: wild-type, anti-FLAG antibody, 2 days [WIP4]; Aspergillus nidulans; ChIP-Seq
To collect samples for ChIP-seq analysis, two-day-old conidia (2×109) of TMK13 were cross-linked with 1% formaldehyde, resuspended in lysis buffer, and homogenized by a mini-beadbeater with 0.5mm zirconia/silica beads. The lysates were then sonicated for five to seven cycles (60 s on, 60 s off) with a sonicator to achieve 150-200 bp size DNA fragments. After centrifugation, the lysates were diluted in the ChIP dilution buffer and applied for ChIP assays using the MAGnify Chromatin Immunoprecipitation System (Invitrogen) follwing the manufacturer's instruction with a modest modification. The diluted chromatin extracts were incubated with 1 μg of mouse monoclonal Anti-FLAG antibody (Sigma-Aldrich). As negative controls, the chromatin extract was reacted with 1 μg of Anti-rabbit IgG. Initial input DNAs before immunoprecipitation were used as positive controls. The enriched DNA fragments were retrieved and used as a template for ChIP-seq. DNA libraries were prepared using TruSeq ChIP library Preparation Kit (Illumina) and sequenced using an Illumina NovaSeq 6000 platform.