alias: GSM3375694
title: GSM3375694: HC_KO_rep4; Mus musculus; RNA-Seq
library_construction_protocol: Whole E9.5 embryos or the left hippocampal CA region were used for RNA extraction by using 700 µL Trizol (Invitrogen) for homogenization and 140 µL chloroform (Sigma), followed by centrifugation at 12,000g for 15 min at 4°C. The upper aqueous phase was transferred to a new tube and 1.5 volumes of 100% ethanol (EtOH) were added. RNA was extracted using Zymo-Spin™ IC columns (Zymo Research). In short, the aqueous phase/ethanol mixture was loaded onto the column, washed with 400 µl 70% EtOH before being treated with RQ1 DNAseI (Promega, 5 µl + 5 µl reaction buffer + 40 µl 70% EtOH) for 15 min at RT. After two washes with 70% EtOH the sample was eluted from the column with DEPC-treated H2O. RNA concentration was measured by using the NanoDrop spectrophotometer (Thermo Scientific). 1 μg (CA) or 500 ng (E9.5) total RNA were used to generate cDNA libraries by following the manufacturer's instructions in the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen). Library quality and size distribution were assessed with the High Sensitivity DNA Analysis Kit for the Bioanalyzer 2100 (Agilent).
identifiers.namespace: GEO
identifiers.id: GSM3375694