Era GTPase of Staphylococcus aureus is a ribosome maturation factor. Era GTPase is involved in the late stages of small (30S) ribosome subunit assembly and regulates formation of the entire 70S ribosome. Study of the structure of the Era and 30S subunit complex from S. aureus will help in studying the processes of ribosome maturation and the mechanisms of protein synthesis regulation in this pathogenic microorganism. This paper presents protocols for obtaining Era GTPase and immature 30S ribosome subunits from S. aureus, as well as a protocol for assembling their complex for further structural studies.