Genome-wide analysis of polyadenylation events in Schmidtea mediterranea
The total RNA amounting to 7ug extracted from both the sexual and asexual worms was subjected to cDNA conversion using OligodT primers from AffinityScript QPCR cDNA Synthesis Kit. The cDNA was fragmented using Covaris. The fragmented DNA was quantified and subsequently utilized for library preparation. End-repair and adapter ligation was carried out according to the protocol (Ion plus fragment library kit, (# 4471252)). The samples were barcoded at this step and then cleaned using Ampure XP beads. The library was amplified using 8 cycles of PCR for enrichment of adapter-ligated fragments. The amplified product was again cleaned using Ampure XP beads. The prepared library was quantified using Qubit and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent). Ion PGM sequencing was performed on this sample.
inStem, NCBS
3P-sex validation