GSM5115828: NP-E-3; Homo sapiens; ncRNA-Seq
Cells are centrifuged and RNA was harvested using Trizol reagent. Illumina TruSeq RNA Sample Prep Kit (Cat#FC-122-1001) was used with 1 ug 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 ~200bp (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.