GSM5034286: RNA-Seq Pre_340_3; Homo sapiens; RNA-Seq
RNA was isolated from cells using the TriPure isolation reagent (Roche) and the Direct-zol Mini Kit (Zymo) following the manufacturers' protocols. Samples were quantified on a NanoDrop ND-1000 instrument. Agarose electrophoresis was used to check the integrality of total RNA samples. 1 ∼ 2 µg total RNA of each sample was taken for RNA-seq library preparation. Briefly, rRNA is removed from the total RNA with a RiboZero Magnetic Gold Kit; The enriched rRNA depleted RNA was used for RNA-seq library preparation using KAPA Stranded RNA-Seq Library Prep Kit (Illumina). The library preparation procedure included: 1) Fragmentation of the RNA molecules; 2) Reverse transcription to synthesis first strand cDNA; 3) Second strand cDNA synthesis incorporating dUTP; 4) End-repair and A-tailing of the double stranded cDNA; 5) Illumina compatible adapter ligation; 6) PCR amplification and purification for the final RNA-seq library. The completed libraries were qualified on Agilent 2100 Bioanalyzer for concentration, fragment size distribution between 400 ∼ 600 bp, and adapter dimer contamination. The amount was determined by absolute quantification qPCR method. The barcoded libraries were mixed in equal amounts and used for sequencing on the instrument.
GEO Accession
GSM5034286