The great plate count anomaly describes the phenomena whereby the yield and diversity of microbes cultured from environmental samples are lower than expected. The difference in expected versus obtained results is due to many factors, including lack of access to environmental nutrients and change in growth conditions. This study examined the effect of various culturing methods on both biodiversity and yield of environmental samples. Culturing techniques utilized included serial dilution plating, filter stamping, filtrate culturing, and semi-solid agar inoculation co-culture. Each technique alters the density of the microbes in the environmental sample either by changing the sample concentration or excluding by size. In addition, co-culturing increases likelihood that growth factors will be produced. Microbial density produced by each method was comparable but differed based on the degree of sample dilution. The diversity of cultured samples, defined by differences in colony morphology, was greatest for the serial dilution plating and co-culturing methods. Results indicate that size exclusion, as well as differences in sample concentration of the various culturing methods, has a great effect on microbe biodiversity in mass culture. Due to the urgent need for novel antibiotics, understanding the effect of mass culturing methods on microbial yield can aid in determining the experimental design for isolating antibiotic-producing microbes from environmental samples.