GSM3464040: S_M microRNA; Schistosoma japonicum; ncRNA-Seq
Worms were recovered by hepatic-portal perfusion with PBS solution at five weeks post infection. The worms were transferred into sterile RPMI 1640 and separated by sex under light microscopy, after which the worms were washed three times with sterile RPMI 1640. Schistosomes were homogenized in a glass homogenizer, and total RNA was isolated using TRIzol reagent according to the manufacturer's instructions (Invitrogen, USA). Small RNA libraries were constructed using NEBNext® Multiplex Small RNA Library Prep Set for Illumina® according to the manufacturer's instructions (New England Biolabs, USA). Briefly, the assay began with 20 μg of total RNA from each sample, the small RNAs (<300 nt) extracted were 5' and 3' extended with adapters, and was transcribed into the first strand cDNA using random hexamer primer, and the products were amplified by PCR. The PCR products purified and size-selected by polyacrylamide gel electrophoresis (PAGE), quantified by Qubit2.0 and examined by Agilent 2100 for insert size,which was followed by quantification of the effective concentration of the constructed library by quantitative PCR. Small RNA sequencing was performed on an Illumina HiseqTM 2500/MiSeq platform (Illumina).