DescriptionHigh-throughput sequencing (HTS) technology enables cost-effective and efficient measurement of the nucleobase sequences of millions of DNA molecules. In order to apply HTS in systematic investigation of metabolic variations and transcription initiation, I developed data analysis pipelines, mathematical methods, and software packages. Using these tools, we have identified sequence determinants in transcription initiation and pausing, which provide mechanistic insights into the interactions between RNA polymerase and DNA template. Also, we have modeled the metabolic variations across different tissues using single-cell RNA-seq data, which provides a novel computational method to infer metabolic variations from the differences in gene expression profiles.