5'P+5'OH-RNA: GsSir2/Ago bound RNA with 5'P and 5'OH
pBADHisA plasmid with the cloned protein GsSir2/Ago from Geobacter sulfurreducens PCA (NP_952414.1) expressed in E.coli DH10B cells together with pCDF-Kn plasmid. GsSir2/Ago with bound RNA was purified as indicated in Zaremba et al., 2021 (doi: https://doi.org/10.1101/2021.12.14.472599) using gentle chemical reagent for cell disruption and buffer solutions with 100mM NaCl. GsSir2/Ago bound RNA was purified using phenol isolation. Purified RNA was treated with T4 PNK (ThermoFisher). Then RNA sample was converted to DNA library using Small RNA-Seq Library Prep Kit (Lexogen cat#052).