GSM7950658: ctrl2-3; Mus musculus; RNA-Seq
Total RNA was extracted from the tissue using TRIzol® Reagent according the manufacturer's instructions (Invitrogen) and genomic DNA was removed using DNase I (TaKara). Then RNA quality was determined by 2100 Bioanalyser (Agilent Technologies) and quantified using the ND-2000 (NanoDrop Technologies). Only high-quality RNA sample(OD260/280=1.8~2.2, OD260/230≥2.0, RIN≥8.0, 28S:18S≥1.0, >10ng) was used to construct sequencing library. Transcriptome library was prepared following Clontech-SMART-SeqTM v4 UltraTM Low Input RNA Kit for sequencing using 10ng of total RNA. Then the synthesized cDNA was conducted into library according to Illumina's library construction protocol. Finally, paired-end RNA-seq sequencing library was sequenced with the Illumina NovaSeq 6000 sequencer (2 × 150bp read length).