Implementation of an autocatalytic signal amplifying protocol in digital droplet assays

This thesis is a first attempt to implement AutoCAR-1 in a digital droplet CRISPR assay, a novel attempt to lower the limit of detection (LoD) in Cas-based DNA detection. AutoCAR-1 amplifies the signal instead of the target by introducing a secondary target, a linearized circular mediator. Difficulties in synthesizing the circular mediator meant that experimental work shifted towards troubleshooting the synthesis protocol. No conclusive solution was found, so we conclude with recommendations for further research into mediator synthesis.