GSM8978452: LVCP, postnatal day 7, n=3; Rattus norvegicus; RNA-Seq
Bulk RNA-seq comprised 5 main timepoints: postnatal day (P) P1, P4, P7, P10 and P60. Rat males were anesthetized, transcardially perfused with sterile RNase free cold saline. Brains were removed and choroid plexuses (CP) from the lateral ventricles were isolated under a stereo microscope and stored at -80ºC. Total RNA extraction was performed using SPLIT RNA Extraction Kit (Lexogen, Viena, Austria). Briefly, the CPs were homogenized in an isolation buffer highly chaotropic. Samples, were then transferred to a phase lock gel column, which contains a gel matrix that, based on the density differences, acts as a barrier between the organic and aqueous phase. Acidic phenol and acidic buffer were added to create a monophasic solution, which allows the separation of genomic DNA into the organic phase. Chloroform was added and the phases were separated by centrifugation for 2min at 12.000xg at 18°C. The RNA, contained in the aqueous phase, was decanted into a new micro tube. The RNA was then precipitated onto a silica column by addition of 1.75x volume of isopropanol. The RNA was further purified by washing the column. Library construction for QuantSeq 3' mRNA