title: GSM9244296: NHEK21, Control; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated using TRI-Reagent (Molecular Research Center, Cincinnati, OH, USA). To obtain global transcriptome data, high-throughput mRNA sequencing analysis was 21 performed on the Illumina (San Diego, CA, USA) sequencing platform. The total RNA sample 22 quality was checked on an Agilent BioAnalyzer (Agilent, Santa Clara, CA, USA) using a 23 Eukaryotic Total RNA Nano Kit according to the manufacturer's protocol. Samples with RNA 24 integrity number > 7 were accepted for the library preparation process. RNA-Seq libraries were 25 prepared from total RNA using the Ultra II RNA Sample Prep kit (New England Biolabs, 26 Ipswich, MA, USA) according to the manufacturer's protocol. Briefly, poly(A) RNAs were 27 captured by oligo dT-conjugated magnetic beads, and then the mRNAs were eluted and 28 fragmented at 94°C. First-strand cDNA was generated by random priming reverse transcription, 29 and after the second-strand synthesis step, double-stranded cDNA was generated. After end 30 repair, A-tailing, and adapter ligation, the adapter-ligated fragments were amplified by 31 enrichment PCR, and finally, sequencing libraries were generated. Sequencing runs were 32 executed on a NextSeq 500 instrument (Illumina) using single-end 75-cycle sequencing. mRNA 33 sequencing (from library preparation to the generation of fastq files) was performed at the 34 Genomic Medicine and Bioinformatic Core Facility, Department of Biochemistry and 35 Molecular Biology, Faculty of Medicine, University of Debrecen (Debrecen, Hungary).
identifiers.namespace: GEO
identifiers.id: GSM9244296_r1