Subject	Parasitology
Specific subject area	Transcriptome analysis of Babesia bovis life stages.
Type of data	Tables Graphic Excel file
How data were acquired	Illumina HiSeq™ 2500 Bio-RAD CFX96™ Real-Time PCR
Data format	Raw Analysed
Parameters for data collection	Babesia bovis blood stages were collected from bovines with acute parasitemia and suspended in TRIzol (Thermo Fisher Scientific, Waltham, MA). Babesia bovis kinetes were collected from infected replete female ticks by extraction via pressurized capillary tubing, pooled, concentrated, and suspended in TRIzol. Total RNA was extracted, and library construction performed according to Illumina TruSeq mRNA library protocols.
Description of data collection	Counts were generated from alignments for each gene using the Subread feature of Counts v1.6.0. Genes without at least 1 read per million mapped reads across all three samples within a group were removed, data was normalized using the TMM method, and analysed for differential expression significance testing using the generalized linear model likelihood ratio test (GLM LRT) method in edgeR v3.20.9. The false discovery rate (FDR) method was employed to correct for multiple testing and genes were termed significantly differentially expressed if their logFold Change (logFC) value was greater than or equal to 1 and the FDR set to 5%. Triplicate RNA samples from B. bovis blood or kinete stages were used for qPCR. Quantitative PCR was performed in triplicate using a Bio-RAD CFX96™ Real-Time PCR Detection System. The transcript level of six genes of interest was normalized by dividing the transcript level of the gene of interest with that of the housekeeping genes. To identify suitable “housekeeping” genes, Excel (Microsoft Office 2013) with NormFinder and the comparative delta-Ct method from the RNA-Seq dataset were used to select ten genes that had no significant fold changes between the blood and kinete stages. The top five housekeeping genes were then used to normalize transcript levels in the qPCR data.
Data source location	The U.S. Department of Agriculture-ARS-Animal Disease Research Unit. Pullman/Washington State United States of America 46.730873 ° N, -117.163475 ° E
Data accessibility	Raw data were deposited at the NCBI Gene Expression Omnibus under accession number GSE144066. Direct URL to data: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE144066.
Related research article	M.W. Ueti, W.C. Johnson, L.S. Kappmeyer, D.R. Herndon, M.R. Mousel, K.E. Reif, N.S. Taus, O.O. Ifeonu, J.C. Silva, C.E. Suarez, K.A. Brayton. Comparative analysis of gene expression between Babesia bovis blood stages and kinetes allowed by improved genome annotation. Int. J. Parasitol. doi: 10.1016/j.ijpara.2020.08.006.