title: GSM4221366: OE_LncR2_0h_2; Homo sapiens; RNA-Seq
library_construction_protocol: For the RNA-seq library, the total RNA was extracted from HK-2 cell samples by using TRIzol Reagent (Ambion) following the manufacturer's  instructions. The total RNA was treated with RQ1 DNase (Promega) to remove DNA. The quality and quantity of the purified RNA were determined by measuring the  absorbance at 260 nm/280 nm (A260/A280) using Smartspec Plus (BioRad). The RNA integrity was further verified by 1.5% agarose gel electrophoresis. For each sample, 5 μg of the total RNA was used to prepare the RNA-seq library. The Ribosomal RNAs were depleted with rRNA (Human/Mouse/Rat) Removal Kit  (Ribo-Zero™, RZH1046, Epicentre, and MGIEasy, 1000005953 for the tissue and HK-2 cell samples, respectively) before they were used for directional RNA-seq library preparation. Purified mRNAs were iron-fragmented at 95 °C, followed by end repair and 5′ adaptor ligation. Then, reverse transcription was performed with RT primer harboring the 3′ adaptor sequence and randomized hexamer. The cDNAs were purified and PCR amplified. PCR products corresponding to 200–500 bps was purified, quantified, and stored at −80 °C until they were used for sequencing. For high-throughput sequencing, the libraries were prepared following the manufacturer's instructions and applied to Illumina HiSeq X Ten system for 150 nt paired-end sequencing.
identifiers.namespace: GEO
identifiers.id: GSM4221366