title: NextSeq 500 sequencing; Bulk RNA-sequencing of mRNA from Sertoli cells isolated by a novel single cell suspension protocol and FACS-sorting
design: Bulk RNA-sequencing of mRNA from Sertoli cells isolated by a novel single cell suspension protocol and FACS-sorting
library_construction_protocol: Sertoli cells were FACS-sorted directly to the RLT lysis buffer. Teste from 21, 28 and 70 days old wild-type males were disected and decapsulated. Seminiferous tubules were disrupted mechanically using a gentle shear force and enzymatically. Obtained single cell suspension was used for staining of Sertoli cells and FACS-sorting. RNA was extracted using a RNeasy180 Plus Micro Kit (Qiagen) according to the manufacturer's protocol. Sequencing libraries were prepared using Smarter Stranded Total RNA-seq Kit v2 Pico Input Mammalian (Takara) according to the manufacturer's protocol.