GSM8880177: NC2; Mus musculus; RNA-Seq
extract protocol：Total RNA was extracted using the Trizol(Invitrogen, CA, USA) library construction protocol：RNA-seq library preparation using a standard mRNA-Seq protocol with oligo(dT) selection and random hexamer priming. Oligo(dT)-attached magnetic beads were used to purify mRNA. Purified mRNA was fragmented into small pieces with fragment buffer at appropriate temperature. Then First-strand cDNA was generated using random hexamer-primed reverse transcription.followed by a second-strand cDNA synthesis and purified using AMPure XP Beads.Afterwards, A-Tailing Mix and RNA Index Adapters were added by incubating to endcDNA repair. The cDNA fragments obtained from previous steps were amplified byPCR, and products were purified by Ampure XP Bead, get the final library.After the library was constructed, Qubit 2.0 was used for preliminaryquantification, and the library was diluted to 1.5 ng/pL. Then Agilent 2100 BioAnalyzerwas used to detect the insert size ofthe library. Qrt-pcr was used to accurately quantify the effective concentration of the library to ensure the quality of the library.