The Germplasm Enhancement of Maize (GEM) Project is a cooperative effort between the USDA-ARS, private industry and public researchers to broaden and enhance the germplasm base of maize. In this program, selected accessions from the Latin American Maize Project, and seven tropical hybrids donated by DeKalb to the GEM Project, were crossed to elite proprietary inbred lines contributed by commercial plant breeding programs. In most cases, the resulting hybrids were crossed to a second commercial inbred line and the resulting 25% exotic hybrids were used as breeding populations for further development. To identify GEM germplasm with value to protein quality breeding programs, we developed a process for evaluating the content of the essential amino acids methionine, lysine and tryptophan in the grain of GEM germplasm that balances the need for multiple-year evaluations with the constantly changing entry list of this germplasm screening program. This process involves annual field trials with common checks. Weak entries are dropped from the trial each year to make room for new entries, while strong entries are retained. Methionine exhibited the most significant variation, followed by lysine and then tryptophan. A number of GEM lines had methionine or lysine levels that were significantly better than Corn Belt checks and some were competitive with high-amino acid checks.