GSM8255524: Nucleus pulposus cells from LSI #1, RNA-seq; Rattus norvegicus; RNA-Seq
Total RNA was isolated from samples using TRIzol reagent. Illumina TruSeq RNA Sample Pre Kit (Cat#FC-122-1001) was used with 1 μg of total RNA for the construction of sequencing libraries. Libraries were prepared according to instructions of Illumina HiSeq X ten accompanying the Ovation RNA-Seq System V2 (M01206 v9). Briefly, RNA was elutated according to the manuscription user guide. Then RNA was fragmented using fragment buffer. cDNA was synthed using dNTPs, DNA polymeraseI and RNaseH. Then cDNA was purified using AMPure XP beads. PCR Was used to amplify with Illumina primers for 15 cycles and library fragments of ~200 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified RNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Illumina HiSeq X ten following the manufacturer's protocols.