title: GSM8583541: NC_2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted via TRIzol (15596-018; Ambion, USA). The RNA was further purified through two rounds of phenol‒chloroform extraction and subsequently treated with RQ1 DNase (M6101; Promega, Madison, WI, USA) to eliminate any DNA contamination. The quality and concentration of the purified RNA were assessed by measuring the absorbance at 260 nm and 280 nm (A260/A280) via an ultrafine spectrophotometer (N50 touch, IMPLEN, Germany). Additionally, the integrity of the RNA was confirmed via 1.0% agarose gel electrophoresis. For each sample, 1 μg of total RNA was treated with RQ1 DNase (M6101, Promega, USA) to eliminate DNA prior to the preparation of a directional RNA-seq library via the VAHTS® Universal V8 RNA-seq Library Prep Kit for Illumina (NR605, Vazyme, China).  mRNAs were isolated via VAHTS mRNA capture beads (N401, Vazyme, China).  The fragmented RNA was then converted into double-stranded cDNA.  After the end repair and A-tailing steps, the DNAs were ligated to VAHTS RNA Multiplex Oligos Set 1 for Illumina (N323, Vazyme, China), followed by amplification of the ligated products.  These products were purified and quantified before being stored at -80°C until sequencing. For high-throughput sequencing, the libraries were prepared for following the manufacturer's instructions and applied to Illumina HiSeq X Ten system for 150 nt paired-end sequencing.
identifiers.namespace: GEO
identifiers.id: GSM8583541_r1