title: GSM3015683: APP_WT_Y_12; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted by homogenizing the left hippocampus in TRIzol (Invitrogen, Carlsbad, CA, USA) using 1ml syringes and 22G/26G needles and purified on mirVana spin columns according to the manufacturer's instructions (Ambion, Austin, TX, USA). RNA purity (260/280 and 260/230 ratios) and integrity was assessed using  Nanodrop ND-1000 (Nanodrop Technologies, Wilmington, DE, USA) and Agilent 2100 Bioanalyzer with High Sensitivity chips (Agilent Technologies, Inc., Santa Clara, CA, USA) and Qubit 3.0 Fluorometer (Life Technologies, Carlsbad, CA, USA), respectively. Library preparation and sequencing was performed at the VIB Nucleomics Core (Leuven, Belgium) using 1 μg of total RNA per sample. Poly-A containing mRNA molecules were purified using Illumina TruSeq® Stranded mRNA Sample Prep Kit (protocol version 15031047 Rev.E) and poly-T oligo-attached magnetic beads. After reverse transcription with random primers, different barcodes were introduced to each sample by ligating a single 'A' base to the 3' ends of the blunt-ended cDNA fragments and multiple indexing adapters and 10 cycles of PCR amplification were performed. 96 libraries were pooled and sequenced in 4 runs using NextSeq 500 High75 Output kits on an Illumina NextSeq 500 instrument (Illumina, San Diego, CA, USA).
attributes.tag: GEO Accession
attributes.value: GSM3015683