show Abstracthide AbstractBacillus thuringiensis (or Bt) is a Gram-positive, soil-dwelling bacterium. RNA-seq data for Bacillus thuringienesis strain CT43 (4CG1) and Bacillus thuringiensis ATCC10792 were obtained using an Illumina High-Seq2000 instrument. Continued improvements and next-generation sequencing cost reductions have made RNA-seq a popular choice for gene expression studies. RNA-seq is revolutionizing the fields of genomics and transcriptomics by enabling a wide range of applications such as strand specific expression detection, transcript fusion, alternative splicing isoforms, SNP detection, genome guided or de novo transcript assemblies. It also enables detection of weakly expressed genes and high genome coverage adds to the attraction as an alternative to microarrays. However, the field of RNA-Seq analysis is still evolving. This study aimed to identify the technical and experimental variations in RNA-seq datasets in order to minimize unwanted variation or account for it in transcriptomic studies.