RNA-seq of Macaca mulatta: adult inferior temporal cortex
We used TruSeq RNA Sample Prep Kit to generate cDNA libraries as per the manufacturer's instructions with the modification of adding spike-in RNAs before the mRNA fragmentation. The spike-in RNAs were synthesized by External RNA Control Consortium (ERCC) by in vitro transcription of de novo DNA sequences or of DNA derived from the B. subtilis or the deep-sea vent microbe M. jannaschii genomes and were a generous gift of Dr. Mark Salit at The National Institute of Standards and Technology (NIST). These were used both to track the brain regions and to normalize expression levels across experiments. Each sample was tagged by adding a pair of spike-in RNAs unique to the region from which the sample was taken. Also, an additional three common spike-ins were added for controlling sequencing error rates, which is not influenced by SNP existence. The TruSeq Kit allows addition of 12 different types of index, thus the multiplexing of samples when they are sequenced. The final product was assessed for its size distribution and concentration using Bioanalyzer DNA 1000 Kit.