NextSeq 500 paired end sequencing; Neurogenesis in Prostate Cancer is driven by Neural Precursors from the Central Nervous System
Neurogenesis in Prostate Cancer is driven by Neural Precursors from the Central Nervous System
Lin-EYFP+ cells were isolated from 8-week-old brain or 16-week-old prostate of DCX-CreERT2/loxp-EYFP/Hi-Myc mice, and were collected in Qiazol (Qiagen). RNA was extracted using the miRNeasy microkit (Qiagen). mRNA libraries were prepared using Smart-Seq v4 Ultra Low Input RNA (Takara, Otsu, Shiga). Briefly, cDNA was synthesized by using the locked nucleic acid (LNA) technology integrated with SMART (Switching Mechanism at 5' End of RNA Template) technology. For each library, post RT-PCR cycle number was adjusted according to the number of cells. Libraries 5 were individually adapted and indexed using the Illumina Nextera XT kit and then, were controlled on the Agilent bioanalyzer (Tapestation 2200, Agilent Technologies). Identical librairies were pooled before sequencing at an average read depth of 70 millions reads per sample. Final libraries were controlled on Tapestation 2200 (Agilent Technologies) and were quantified with fluorimetric intercalant.