title: GSM2829243: Day13; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: Total RNA from synchronized adult worms was extracted by using TRI-Solution (Bio Science Technology). Sequencing libraries were manually generated for high-throughput sequencing. Poly(A) mRNA was purified using Oligotex Direct mRNA Mini Kit (Qiagen). Enriched mRNA was fragmented by incubating in 5X RNA Fragmentation Buffer (200 mM Tris-acetate (pH 8.1), 150 mM MgOAc, and 500 mM KOAc) at 82 °C for 5 min. cDNA was synthesized by using SuperScript III RT (Invitrogen) and random hexamers with incubation for 30 min at 55 °C. Approximately 500 ng of cDNA of each sample was repaired using the End-It DNA End-Repair Kit (Epicentre Biotechnologies), followed by addition of 3' A base for adaptor ligation. Overnight ligation of polished cDNA with Genomic DNA Adaptors (Illumina), amplification for 18 cycles, and isolation of about 200~500 bp from agarose gel were performed according to the manufacturer's instructions.
identifiers.namespace: GEO
identifiers.id: GSM2829243