Loss of nitrogen (N) from dairy manure during storage is an issue of economic, environmental, and social concern for farming communities. The lost N 1) decreases the value of manure as a fertilizer and is an economic loss because supplemental inorganic N fertilizer is purchased to meet N needs on farms; 2) produces the potential pollution for water and air systems, thereby damaging the associated ecosystems; 3) causes challenges to human health. Thus, it is vital to manage and use N in an efficient and eco-friendly manner. N mineralization is a pathway in the N cycle, which converts organic N to inorganic N that is more susceptible to loss. The objective of this study was to conduct lab-scale experiments to assess the effects of temperature, manure solids content, using manure seed and autoclave sterilization operation at the start of storage, and storage time on the N mineralization and the associated microbial community during the storage of liquid dairy manure. Manure scrapped from the barn floor of a commercial dairy farm and diluted to make experimental stocks with high (46 to 78 g/L) and low (19 to 36 g/L) total solids (TS), to simulate what is typically transported to the manure storage pit was used. The manure was incubated in the laboratory at three temperatures (10, 20, and 30°C) for two storage periods (60 and 180 days). Manure samples were taken at different storage time for analyses. The results showed that temperature and using sterilization operation at the start of storage had significant effects on N mineralization for both storage periods (p < 0.05). The highest N mineralization rate occurred at 30℃, which rate constant (k) was 0.096 week-1. While, the lowest N mineralization occurred at 10℃, and its corresponding k was 0.013 week-1. The concentrations of mineralized N (Nm) with non-sterilized (R) manure were significantly higher than that with sterilized (R0) manure (p < 0.05). Compared to that with high TS (H) manure, the concentrations of Nm were significantly higher with low TS (L) manure after 180-d storage (p < 0.05). Raw manure augmented with manure seed (MS) had significantly higher Nm than the manure seed only (SO) (p < 0.05). In order to investigate the changes of microbial community in manure, samples were collected on days 0, 30, 90, and 180 for the 180-d storage experiment, and days 0, 30, and 60 for the 60-d storage experiment, and then manure DNA under different condition was successfully extracted from collected samples and used for 16S rRNA sequencing. This study provided a more comprehensive understanding of the impact factors for manure storage, and was expected to clarify the relationship between N mineralization and the associated microbial community.