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Date of Award
Molecular Biology and Genetics
Complex traits and diseases are associated with variants in non-coding regions of the genome. These variants have shown to alter phenotypes as well. Characterizing these variants is therefore important and more recently has been made possible with high throughput reporter assays like STARR-seq. A novel method developed in our lab, BIT-STARR-seq based on STARR-seq is able to further identify allele-specific expression in synthesized oligos representing regulatory regions in the DNA while using unique molecular identifiers to account for PCR duplicates. In this study, we optimize the library preparation protocol for BiT-STARR-seq for the HUVEC cell line using the STARR-seq human plasmid. We test the efficiency of the protocol and troubleshoot problems we came across. Finally, we sequence 5 replicates of the newly generated library and quantify their presence in transformed cells.
Divgikar, Advait, "High Throughput Reporter Assays To Characterize Regulatory Variants" (2017). Wayne State University Theses. 615.