Effect of seed mismatches on target cleavage by MILI, HILI and EfPiwi

To measure the effect of seed mismatches on target cleavage by MILI, HILI and EfPiwi, sixty-three target RNAs with different combinations of mismatches to g2-g8 of piRNA sequence were combined in equimolar amounts and incubated with MILI-piRNA, HILI-piRNA and EfPiwi-piRNA complexes, respectively. The corresponding cleavage products were analyzed by deep sequencing.