A novel aqueous two-phase system (ATPS) only containing Gemini surfactant ethanediyl-1,2-bis(dodecyldimethylammonium bromide) and an inorganic salt sodium bromide was designed, and the physicochemical properties of the ATPS were investigated systematically. The results have shown that the coexisting two phases, one surfactant-rich and the other salt-rich, are stable and the phase behavior, volume ratio, and extraction efficiency of ATPS are strongly influenced by contents of surfactant and salt. The novel ATPS here investigated have potential application in partitioning and analysis of biomaterials. Compared with other complicated surfactant-based ATPS, the surfactant concentration for the formation of ATPS is much lower, which implies that the studied system is potentially more economical for purification and separation of biomaterials. Meanwhile, the surfactant can be recycled by altering salt concentration in ATPS. Lyotropic liquid crystal was found in the bottom phase besides micelles under different conditions. The salting-out effect may be the driving force of ATPS formation for the studied "living polymer"/salt system. However, the coexistence of micelles with different microstructures is the necessary condition of phase separation. The current studies not only present a new ATPS for partitioning and analysis of biomaterials but also have a great significance on rational use of Gemini surfactants.