Next Generation Sequencing Facilitating Quantitative Analysis of Wild Type (Kcnj11fl/fl, WT) and β-cell Specific Kcnj11 -/- (βKcnj11 -/-, bKO) Mouse Islets' Transcriptomes

β-cell Specific Kcnj11 -/- mice and their islets exhibited a hirtherto unkown switch in the major signaling pathway of G-protein coulpled receptors from Gs to Gq, and overall enhanced Gq signaling. That the phenomeno was also observed in their isolated islets, and that the genetic manipulation of the mice was only in the pancreatic beta-cells, we reasoned that peharps knockout of Kcnj11 in beta cells could result to a general change in the expression of genes related to G-protein coupled receptor signaling. Since New-Generation Sequencing (NGS) has revolutionized cellular based pathway analysis, we extracted total RNA from WT and bKO islets, performed NGS and compared global changes in the expression of genes between WT and bKO islets. WT and bKO RNA samples, in triplicates, were processed as fragments per kilobase of transcript per million mapped reads (FPKM), and determined by the RNA-sequencing (RNA-seq) service provider Macrogen, who also mapped trimmed reads to the Mus musculus reference genome using AnnoOnly(-G) Transcriptome Resequencing NGS Advanced and then performed transcript assembly using Cufflinks-G. Data were log transformed and quantile normalization was performed with Pre-process Core’s R library by Macrogen. Samples with at least one FPKM value of 0 were removed from consideration