title: GSM8877427: rde-4(ne301), young adults, RNA-Seq, biol rep 1; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: Populations of young adults were collected and washed in M9 and added to Trizol Reagent (Invitrogen). Worms in Trizol were lysed by freeze-thawing from -80 °C to 4 °C three times. Total RNA was purified from the lysate using the miRNeasy kit (Qiagen) according to manufacturer instructions, including the optional Buffer RWT wash step. 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. Library construction was performed by Novogene Corporation.
identifiers.namespace: GEO
identifiers.id: GSM8877427_r1