title: GSM1213668: NP214 (mRNA-Seq); Rattus norvegicus; RNA-Seq
library_construction_protocol: For tissue preparation, ether anesthetized animals were decapitated, the sculls opened and cortex removed for preparation of dorsal hippocampus. Five mm thick hippocampus slices were prepared along the septo-temporal axis and collected in ice cold PBS, thereby, removing blood contamination. Hippocampi were snap frozen in liquid nitrogen and stored at -80oC until further use. Each probe was individually processed and not pooled with other samples. For downstream applications, hippocampus was homogenized in ice cold 1x PBS and divided into equal volumes for DNA and RNA extraction. 10 mg rat hippocampal tissue was used for total RNA extraction using the Trizol method, followed by DNase digestion. RNA quality was verified on the Shimadzu MultiNA capillary electrophoresis system (Shimadzu). Poly-A mRNA was isolated with Dynabead Oligo(dT) enrichment (Invitrogen). Sequencing libraries were prepared using the NEBNext® mRNA Library Prep Reagent Set for Illumina® (New England Biolabs). Libraries were quantified on MultiNA system and sequenced at a concentration of 12 pM on the Genome Analyzer IIx (Illumina Inc, CA, USA) using the TruSeq version 5 cluster kit (cBot) and SBS kit (36 cycles).
attributes.tag: GEO Accession
attributes.value: GSM1213668