title: GSM7984502: iPSC_1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from the tissue using TRIzol® Reagent according the manufacturer's instructions. Then RNA quality was determined by 5300 Bioanalyser (Agilent) and quantified using the ND-2000 (NanoDrop) RNA purification,  reverse  transcription,  library  construction  and  sequencing  were performed  at  Shanghai Majorbio  Bio-pharm  Biotechnology  Co.,  Ltd.  (Shanghai,  China)  according  to  the  manufacturer's  instructions (Illumina, San Diego, CA). The RNA-seq transcriptome librariy was prepared following Illumina® Stranded  mRNA Prep, Ligation from Illumina (San Diego, CA) using  1μg of total RNA.  Shortly, messenger RNA was isolated according to polyA selection method by oligo(dT) beads and then fragmented by fragmentation buffer firstly. Secondly double-stranded cDNA was synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, CA) with random hexamer primers (Illumina). Then the synthesized cDNA was subjected to end-repair, phosphorylation and  'A' base addition according to Illumina's library construction protocol. Libraries were size selected for cDNA target fragments of 300 bp on 2% Low Range Ultra Agarose followed by PCR amplified using Phusion  DNA  polymerase  (NEB)  for  15  PCR  cycles.  After  quantified  by  Qubit  4.0,  paired-end  RNA-seq sequencing library was sequenced with the NovaSeq 6000 sequencer (2 × 150bp read length).