Incorrect deployment of RFID readers occurs reader-to-reader interferences in many applications using RFID technologies. Reader-to-reader interference occurs when a reader transmits a signal that interferes with the operation of another reader, thus preventing the second reader from communicating with tags in its interrogation zone. Interference detected by one reader and caused by another reader is referred to as a reader collision. In RFID systems, the reader collision problem is considered to be the bottleneck for the system throughput and reading efficiency. In this paper, we propose a novel RFID reader anti-collision algorithm based on evolutionary algorithm(EA). First, we analyze characteristics of RFID antennas and build database. Also, we propose EA encoding algorithm, fitness algorithm and genetic operators to deploy antennas efficiently. To show superiority of our proposed algorithm, we simulated our proposed algorithm. In the result, our proposed algorithm obtains 95.45% coverage rate and 10.29% interference rate after about 100 generations.