PRJNA516025
Mus musculus breed:C57BL/6J Genome sequencing
RNA sequencing from the medial Prefrontal Cortex (mPFC) tissue from 3 months old and aged 18 months old mice. Mice were further separated in a cohort that has been subjected to a 10 minute locomotion test followed by an active avoidance learning task prior to tissue collection (young =3 and aged =3 mice) and a cohort that was behaviorally naive (young =3 and aged =3 mice).Total RNA was extracted from mPFC, using the AllPrep DNA/RNA/miRNA Universal Kit (QIAGEN, Hilden, Germany) with an on-column DNAse treatment step (to remove genomic DNA), following the manufacturer's instructions. Isolated RNA quality (integrity) was assessed on an Agilent RNA 6000 Nano Chip (Agilent Technologies, Santa Clara, CA, USA) using the Agilent 2100 BioAnalyser (Agilent Technologies, Santa Clara, CA, USA), following the BioAnalyser instructions.Libraries were prepared with TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero (Illumina, San Diego, CA, USA). Indexed libraries were quantified with Agilent Technologies 2100 Bioanalyzer with the DNA 1000 kit (Agilent Technologies) and KAPA Library Quantification Kit for Illumina sequencing platforms (Roche Applied Science, Basel, Switzerland). Libraries were diluted and six individual libraries were pooled to a final concentration of 2nM. Pooled libraries were quantitated with KAPA Library Quantification Kit for Illumina sequencing platforms (Roche Applied Science). RNA sequencing was performed using Illumina HiSeq 2500 System with 100bp paired-end sequencing.