Real-time qPCR Analysis of S. mansoni miRNA

Real-time quantitative PCRs were performed across two life cycle stages of S. mansoni representing juvenile (28 days post-infection, d.p.i.) and adult worms (49 d.p.i.) using TaqMan Small RNA Assays (Supplementary S1 Text) purchased from Applied Biosystems (Life Technologies, UK). Reverse transcription and quantitative PCR experiments were performed according to manufacturer’s instructions in a StepOnePlus RT-qPCR machine (Applied Biosystems, Life Technologies, UK). S. mansoni U6 was used as the endogenous control [20 (link)] (gene entry sma.U6.1.1 in http://www.genedb.org/Homepage/Smansoni) and fold changes to miRNA expression were estimated using the delta-delta-Ct method [67 (link)].