Updated project metadata. Spectral libraries generated by data-dependent acquisition (DDA) are a useful tool for the analysis of data created by data-independent acquisition (DIA) in mass spectrometry. The quality of DIA analysis is dependent on the quality of the spectral library. We used retinal rat tissue to create a spectral library of rats retina proteome. This data set may therefore be valuable for the future analysis of retinal proteins and research projects that focus on ocular disorders or function.