title: GSM8951022: M2; Mus musculus; RNA-Seq
library_construction_protocol: RNA Extraction: Total RNA was extracted using the Trizol reagent kit (Invitrogen, Carlsbad, CA, USA). The RNA quality was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and RNase free agarose gel electrophoresis.  mRNA Enrichment: Eukaryotic mRNA was enriched using Oligo(dT) beads for eukaryotic samples or Ribo-ZeroTMMagnetic Kit (Epicentre, Madison, WI, USA) for prokaryotic samples.  Fragmentation and cDNA Synthesis: mRNA was fragmented and reversed transcribed into cDNA using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB #7530, New England Biolabs, Ipswich, MA, USA).  Library Construction and Sequencing: The cDNA was end repaired, an A base was added, and Illumina sequencing adapters were ligated. The library was then amplified by PCR and sequenced using Illumina Novaseq6000 by Gene Denovo Biotechnology Co. (Guangzhou, China). The cDNA fragments were purified using AMPure XP Beads (1.0X), end-repaired, A-tailed, and ligated with Illumina adapters.  The final library was amplified by PCR.