Zucchini Yellow Mosaic Virus (ZYMV) is a plant virus with an RNA genome found within the Potyviridae family. This virus causes chlorosis, or yellowing of the leaves, in infected plants. Plants infected with ZYMV often yield unmarketable fruits, resulting in economic losses across the United States and the world. Primers were designed to amplify a segment of the viral genome using RT-PCR. These primers contained EcoRI and XbaI restriction sites, allowing for insertion of the replicated insert into a plasmid vector with those same sites. The plasmid, pUC57, was digested with EcoRI and XbaI and subjected to phosphatase treatment, to ensure the plasmid would not self-ligate. The insert and plasmid were ligated together and transformed into competent E. coli cells. These cells were plated on petri plates with LB, X-gal, and IPTG to allow for blue-white colony screening. Colonies appearing white indicate the transformation of the plasmid and white colonies will be screened for the insert by colony PCR and subsequent quantitative PCR (qPCR).