GSM5212259: 3mo_1mpi_AD_5 [X6089.1mpi.plus]; Mus musculus; RNA-Seq
Qiagen RNeasy Mini Kit Mature messenger RNA was isolated using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) followed by fragmentation and priming with NEBNext First Strand Synthesis Reaction buffer and NEBNext Random Primers. First and second strand synthesis were completed using ProtoScript II Reverse Transcriptase and Second Strand Synthesis Enzyme Mix (NEB), respectively. Double-stranded cDNA was purified with AxyPrep Mag PCR Clean-up (Axygen) and End Prep Enzyme Mix was used to repair ends and A-tail followed by addition of adaptors with T-A ligation (NEB). AxyPrep Mag PCR Clean-up was used for selection of ~360bp fragments and these were amplified with 11 cycles of PCR using P5 and P7 primers. Libraries were purified with AxyPrep Mag PCR Clean-up and quantified using and Agilent 2100 Bioanalyzer and Qubit 2.0 Fluorometer (Invitrogen).