Figure 2. TtAgo guides are 5′-phosphorylated DNA molecules.

a, Schematic representation of TtAgo and TtAgoDM proteins used for all experiments (N, PAZ, MID, and PIWI are structural domains, L1 and L2 are linkers⁸). The amino-terminal Strep(II)-tag is indicated as a black square. b, Co-purified nucleic acids from TtAgo and TtAgoDM are labelled with [γ-³²P]ATP after phosphate exchange by PNK from bacteriophage T4, and treated with enzymes as indicated. M, custom ssDNA marker; nt, nucleotides. c, Length distribution of unique ssDNA sequences co-purified with TtAgo. d, Nucleotide composition of unique ssDNA sequences co-purified with TtAgo. e, Unique reads of TtAgo co-purified ssDNA molecules mapped on the TtAgo expression vector pWUR702.