RNA (10 µg) isolated from S. japonicum schistosomula were separated on a 15% denaturing PAGE gel and transferred to a nylon membrane. Membranes were incubated with biotin-labeled probes complementary to candidate miRNA sequences (sja-miR-1a, sja-miR-1b, sja-miR-7, sja-miR-71b, sja-miR-71a, sja-miR-87, sja-miR-124, sja-miR-281, sja-miR-2b, sja-miR-2a, sja-miR-307, sja-miR-31, sja-miR-133, sja-miR-310, sja-miR-8, sja-miR-10, sja-miR-36, sja-miR-61, sja-miR-277, sja-miR-219, sja-miR-190, sja-miR-candidate-03). The detection of hybridization at 20 nucleotides indicates the expression of the corresponding sequences. Northern analysis with several of the probes do not reveal hybridization at 20nt indicating either that they are expressed at too low a level to be detected or that they are not expressed at all. M1: biotin-labeled marker; M2: 10bp DNA marker.