GSM4675889: Rat-Brain-RNaseR; Rattus norvegicus; RNA-Seq
RNA purchase/extraction: Human Adult Normal Brain Total RNA (R1234035-50), Human Adult Normal Cerebellum Total RNA (R1234039-50), Rat Brain Total RNA (R1434035-50), Monkey(Rh) Brain (R1534035-50) Total RNA was obtained by BioChain. Human Blood Total RNA (HR-705) was obtained from Zyagen. One C57BL/6 male and one female mouse were sacrificed at p21, different brain regions were harvested (cerebellum, cortex, olfactory bulb, brain stem and thalamus) by the Nelson's lab at Brandeis University. The RNA was extracted with TRI reagent (Sigma, Aldrich), followed by Zymo RNA cleaning and concentrator -25 Kit (Zymo), according to the manufacturers' instruction. Recovered RNA was used to prepare a total RNA library, adapting the protocol from Shishkin et al., Nature 2015. Specifically, we prepared one library for the RNaseR treated samples and one for the Mock ones. Briefly, RNA was fragmented for 3 min at 92°C in FAstAP 2X buffer (ThermoFisher). Subsequently, samples were dephosphorylated with FastAP enzyme (ThermoFisher), the first linker was ligated, and samples belonging to the same library were pulled together. Retro-transcription was performed using Affinity Script (Agilent). After second ligation, we performed PCR to add the external barcode. RNaseR vs. Mock ribodepleted total RNA library. The samples were sequenced by Novogene Corporation Inc. (8801 Folsom BLVD, Suite 290, Sacramento, CA 95826) - HiSeq 1% PhiX.