Instrument: Illumina NovaSeq 6000
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: SINGLE
Construction protocol: Genomic DNA was extracted with Promega wizard genomic kit according to the manufacturers protocols for gram negative bacteria. 6XHIS-tagged DNA-binding response regulator proteins were heterologously expressed in E. coli, purified with metal affinity resin, and mixed with 60 uL DNA binding buffer before elution. DNA binding buffer contained 0.4 ng/uL sheared adapter-ligated Pseudomonas genomic DNA, 10 mM imidazole, 0.1% Tween20 with or without 50 mM acetyl phosphate in TBS buffer pH 7.4. To enrich for specific protein-DNA interactions, resin bound protein-DNA was washed in a low concentration of imidazole before elution in a high concentration of imidazole. DNA was then amplified with index primers and sequenced with Illumina NovaSeq 6000. DNA libraries were prepared for sequencing using a protocol from Chiniquy et al., BMC Genomics, 2020.