title: GSM1479217: mBrain_RNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from brain using TRIzol Poly-A+ RNA was isolated with OligodT25 dynabeads (Invitrogen, #610-02) from 2 ug total RNA of each sample. Reverse transcription was performed with random hexamers and Superscript III reverse transcriptase (Invitrogen), followed by the second strand synthesis with E. coli DNA PolI (Invitrogen). Double stranded cDNA was purified with Qiaquick PCR purification kit (Qiagen), and sheared with a nebulizer (Invitrogen) to 100-500 bp fragments. These fragments were then end-repaired using End-It Repair Kit (Epicenter), tailed with an A using Klenow exo minus (NEB, #M0212), and ligated to custom adapters with T4 DNA ligase (Enzymatics, #L603-HC-L). Fragments of 200±20 bp were size-selected and subjected to ligation-mediated PCR amplification (LM-PCR), using Phusion DNA polymerase (NEB, #M0530).
attributes.tag: GEO Accession
attributes.value: GSM1479217