GSM7527883: 1383D2-day13+7; Homo sapiens; RNA-Seq
Total RNA were extracted using RNeasy according to the manufacture's protocol. For RNA sequencing, total RNA was extracted as described above. The samples preserved at −80°C were shipped and analyzed by DNAFORM. The quality of total RNA was evaluated by a Bioanalyzer (Agilent) to ensure over 8.0 RIN (RNA integrity number) or electrophoresis waveform. Double-stranded cDNA libraries (RNA-seq libraries) were prepared using a SMART Seq Stranded Kit (Clontech) according to the manufacturer's protocols. RNA-seq libraries were sequenced using paired-end reads (50 nt of read 1 and 25 nt of read 2) on a NextSeq 500 (Illumina). Obtained reads were mapped to the human GRCh38 genome analyzed by STAR (version 2.7.3a). Annotated reads were counted using featureCounts (version2.0.1) and RSEM (version 1.3.1). FPKM values were calculated from mapped reads by normalizing to total counts.