title: GSM4802317: RNA-seq_miR630_2; Homo sapiens; RNA-Seq
library_construction_protocol: For each sample, 5μg of total RNA was used for RNA-seq library preparation. Polyadenylated mRNAs were purified and concentrated with oligo (dT)-conjugated magnetic beads (invitrogen) before used for directional RNA-seq library preparation. Purified mRNAs were iron fragmented at 95℃ followed by end repair and 5' adaptor ligation. Then reverse transcription was performed with RT primer harboring 3' adaptor sequence and randomized hexamer. The cDNAs were purified and amplified and PCR products corresponding to 200-500 bps were purified, quantified and stored at -80℃ until used for sequencing. For high-throughput sequencing, the libraries were prepared following the manufacture's instructions and applied to Illumina NextSeq500 or Illumina Hiseq X Ten system with 150x2 paired-end type (ABLife Inc., Wuhan, China).