design: Wound samples (blastema and post-blastema) were obtained at 0h of regeneration after amputation on wnt1 RNAi animals. Three biological replicates were used per time point; each biological replicate was composed of eight animal fragments. At the corresponding time point animals were placed in a Petri dish with cold 1% HCl (diluted in water) for 2 and then transferred to a new Petri dish with cold PBS 1X. Two washes were performed with cold PBS 1X and animals were transferred to cold RNAlater for 20 placed in ice. Then, planarians were amputated in a Peltier Cell with a clean blade, to obtain the blastemas and post-blastemas. Fragments were washed with RNAlater (Invitrogen) and 50% RNAlater/Trizol; liquids were removed and 100ul of Trizol reagent (Invitrogen) was added, samples were then treated with DNase I (Roche). RNA-Seq libraries were prepared with KAPA Stranded mRNA-Seq Illumina Kit following the instructions provided by the manufacturer (Roche-Kapa Biosystems). Briefly, 500ng of total RNA was used for the poly-A fraction enrichment with oligo-dT magnetic beads, following the mRNA fragmentation. The strand specificity was achieved during the second strand synthesis performed in the presence of dUTP instead of dTTP. The blunt-ended double stranded cDNA was 3adenylated and Illumina platform compatible adaptors with unique dual indexes and unique molecular identifiers (Integrated DNA Technologies) were ligated. The ligation product was enriched with 15 PCR cycles and the final library was validated on an Agilent 2100 Bioanalyzer with the DNA 7500 assay. The libraries were sequenced on HiSeq 4000 (Illumina) with a read length of 2x76bp using HiSeq 4000 SBS kit (Illumina) and HiSeq 4000 PE Cluster kit (Illumina), following the manufacturers protocol for dual indexing. Image analysis, base calling and quality scoring of the run were processed using the manufacturers software Real Time Analysis (RTA 2.7.7).