GSM6996049: dot-1.1(knu339); ced-3(n1286) embryos biol rep 3; Caenorhabditis elegans; ssRNA-seq
miRNeasy kit (Qiagen) following standard procedures for total RNA extraction, including the optional wash with RWT buffer Strand-specific libraries were generated using NEBNextUltra RNA Library Prep Kit for Illumina (NEB, USA) following manufacturer's recommendations. Ribosomal RNA was depleted. In order to select cDNA fragments of preferentially 250-300 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA). Then USER Enzyme (NEB, USA) was used to digest the second strand with size selected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR. Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.