Thatch-mat and organic matter (OM) accumulation near the putting green soil surface impacts soil physical properties and turf performance. Excessive thatch and OM can encumber infiltration of water and oxygen into the soil profile and slow drainage of excess water away from the putting surface. Proper sampling of thatch-mat depths and OM contents is vital for management of putting green turf; therefore, a study was performed in Knoxville, TN, to derive proper sampling procedures of these important parameters using ‘TifEagle’ and ‘Champion’ bermudagrass (Cynodon dactylon × C. transvaalensis), ‘SeaDwarf’ seashore paspalum (Paspalum vaginatum), and ‘Diamond’ zoysiagrass (Zoysia matrella). ‘TifEagle’ and ‘Champion’ accumulated thatch-mat to a greater depth than ‘SeaDwarf’ and ‘Diamond’. However, ‘SeaDwarf’ had a higher OM content than ‘Diamond’ and both had higher OM contents than ‘TifEagle’ and ‘Champion’. Data generated from sampling procedures indicate that previous studies often undersampled plots for thatch-mat depth; however, previous sampling procedures have not traditionally undersampled plots for OM. Data in this study provide a range of confidence and minimum detectable difference levels which may allow future researchers to more accurately sample ‘TifEagle’, ‘Champion’, ‘SeaDwarf’, and ‘Diamond’ putting green plots for thatch-mat depth and OM content.