Template Switching 2.0: A Microfluidic Approach
Faculty Mentor: Ángel Islas
Template switching is a phenomenon in which an enzyme synthesizes DNA across two non-continuous, juxtaposed DNA templates through microhomology between the two strands . Template switching activity and substrate preference have been previously detailed by Garcia et. al using a highly sensitive template switching assay designed in the Islas lab. Continued work using this assay revealed other enzymes that have template-switching activity, including the p66 subunit of HIV reverse transcriptase, further validating the assay’s capability as a standard for studying template switching. This assay, however, is time, labor and reagent intensive. The advent of microfluidics (micro-volume experiments on designed micro-platforms) for biological purposes has opened the doors for increasingly rapid, sensitive biological assays. This project aims to move the existing template switching assay to a micro-platform for rapid, cheaper analysis and ease of experimental execution with comparable sensitivities to the existing template switching assay.
 García, Patty B., Nicole L. Robledo, and Ángel L. Islas. “Analysis of Non-Template-Directed Nucleotide Addition and Template Switching by DNA Polymerase.” Biochemistry 43.51: 16515-6524 (2004).