GSM8683485: mouse circRNA Brain WT 1646 batch 1; Mus musculus; ncRNA-Seq
Tissues were extracted and frozen on dry ice. A bead mill (Retsch, Düsseldorf, Germany) was used to pulverize the tissue. RNA was subsequently extracted from the tissue powder using a phenol-based extraction protocol (PeqGold TRIFAST, VWR) according to the manufacturer's protocol. RNA integrity was visualized using a Bioanalyzer (Agilent, Santa Clara, CA, USA) 5 µg total RNA was rRNA depleted using hybridization based protocol (siTOOLS Biotech, Martinsried, Germany). The RNA was subsequently polyadenylated and digested with Rnase R for digestion of linear fragments. After purification the remaining RNA was fragmented and used for library construction using the NEBNext Ultra II RNA Library Kit. Libraries were sequenced using the Novaseq 6000 platform (Illumina, San Diego, CA, USA)