As a bi-nucleated eukaryote, Tetrahymena thermophila has two distinct nuclear cycles within each vegetative cell cycle. Rather than a mononucleate G1-S-G2-Mitosis cell cycle, the somatic nuclear cycle of Tetrahymena progresses through G1-Smac-G2-Amitosis, while the germline nuclear cycle is Smic-G2-M-Smic. Furthermore, in contrast to mononucleated organisms, mitosis is not temporally coupled to cytokinesis. Gene expression data could reveal transcriptional programs critical to processes during the cell cycle. By use of centrifugal elutriation to synchronize the vegetative cell cycle, along with RNAseq and EdU labeling, we have begun to elucidate the transcriptional program across the vegetative cell cycle. At 30-minute intervals, we have deployed RNAseq to identify all genes that encode cell cycle regulated transcripts. We have also subjected elutriated cells to EdU labeling to assess mic and mac DNA synthesis, and in conjunction with other cytological markers have assessed synchrony and assign time points to stages of the cell cycle. What we have found is that of the ~27000 annotated genes (TGD.org), nearly 4500 (16%) exhibit a dynamic regulation in mRNA levels of a two-fold change above background. Of those 4500 genes, 2,166 (48%) map to an associated gene ontology (GO) term. At our most stringent criteria for cycling – above twice the median expression of all genes (around 50 counts after normalization) and at least a 4-fold change in expression between 2 time points – we find roughly 400 genes of which 139 (35%) map to associated GO terms. 69 (49%) of the GO terms assigned to our most stringent criteria fall in pathways involved in DNA replication, chromosome organization and microtubule anchoring. Examples of genes within these pathways that demonstrate a 4-fold change in expression include TPB1, a PiggyBack-like protein, DNA licensing factor MCM2, and separase protein ESP1. In regards to temporal regulation of mRNA production, we have found that mRNA levels of replicative genes and certain histone modifiers are up-regulated coordinately at the G1/S border and during S phase of the cell cycle. As for genes involved in cell cycle progression – better known as cyclin genes – we see that 13 of the 26 show dramatic regulation in mRNA levels and meet our criteria for cycling. Some cyclin profiles may suggest their involvement mic –or mac-specific events, warranting further investigation. The wide breath of pathways and genes covered by our data suggest its usefulness to the research community.