A highly anticipated update of the previous edition, In Vitro Transcription and Translation Protocols, Second Edition, provides molecular biology laboratories with the most powerful techniques for exploiting in vitro transcription and translation systems.In Section One: Technologies, authors discuss the use of alternative energy systems for ATP regeneration, the efficient in vitro production of integral membrane proteins, and the high-throughput production of protein libraries using the innovative single-molecule PRC-linked in vitro expression (SIMPLEX) technology. In Section Two: Applications, authors present in vitro translation of amplified protease gene from HIV patients to select proper anti-protease drug therapy, in vitro translation of genes from human patients to diagnose biologically relevant gene mutations, use of SIMPLEX technology to select enantioselective lipases from libraries of randomly produced lipase mutants, and the in vitro transcription and translation to identify proteins in 2D-maps.