GSM3723743: NM ProQ FLAG  +crosslink replicate 2; Neisseria meningitidis 8013; RIP-Seq
Bacteria were lysed and the FLAG-tagged proteins was immunoprecipitated using a monoclonal anti-FLAG antibody. The samples were treated with benzonase nuclease, calf intestine phophorylase, and polynucleotide kinase in the presence of radioactive gamma-ATP. Samples were separated with SDS-PAGE and followed by transfer to nitrocellulose membranes. Radioactively labelled RNA-protein complexes were eluted from membranes and treated with Proteinase K. RNA was purified with phenol:chloroform extraction and ethanol precipitation. Purified RNA was used as input for library preparation using the NEB Next Small RNA Library kit according to the manufacturer's instructions. Purified RNA was used as input for library preparation using the NEB Next Small RNA Library kit according to the manufacturer's instructions.