GSM3732616: T4-Punch2; Macaca fascicularis; RNA-Seq
Brain punches were flash frozen in liquid nitrogen at time of collection. RNA was isolated using Monarch® Total RNA Miniprep Kit (NEB, T2010S) following manufacturer protocol. Briefly, DNA/RNA protection solution was added to frozen tissue and allowed to thaw at room temperature. Tissue in solution was homogenized by passing through 27G needle and inubated for 30 minutes at 55C with proteinase K. Lysate was then passed through gDNA removal column, bound to RNA column, subjected to on-column DNase treatment, and eluted with nuclease free water. Library preparation was performed using TruSeq® Stranded mRNA Library Prep (Illumina, 20020594) following manufacturer protocol. Briefly, mRNA was pulled isolated from total RNA via poly-A selection beads. cDNA synthesis was performed using ProtoScript® II Reverse Transcriptase (NEB, M0368L) followed by second strand synthesis. 3' ends were adenylated and indexed adaptors ligated. Size selection was done using Ampure XP beads (Beckman Coulter, A63881).
GEO Accession
GSM3732616