The need for novel antibiotics is greater now than perhaps any time since the pre-antibiotic era. Indeed, the recent collapse of many pharmaceutical antibacterial groups, combined with the emergence of hypervirulent and pan-antibiotic-resistant bacteria has severely compromised infection treatment options and led to dramatic increases in the incidence and severity of bacterial infections. This collection of reviews and laboratory protocols gives the reader an introduction to the causes of antibiotic resistance, the bacterial strains that pose the largest danger to humans (i.e., streptococci, pneumococci and enterococci) and the antimicrobial agents used to combat infections with these organisms. Some new avenues that are being investigated for antibiotic development are also discussed. Such developments include the discovery of agents that inhibit bacterial RNA degradation, the bacterial ribosome, and structure-based approaches to antibiotic drug discovery. Two laboratory protocols are provided to illustrate different strategies for discovering new antibiotics. One is a bacterial growth inhibition assay to identify inhibitors of bacterial growth that specifically target conditionally essential enzymes in the pathway of interest. The other protocol is used to identify inhibitors of bacterial cell-to-cell signaling. This e-book — a curated collection from eLS, WIREs, and Current Protocols — offers a fantastic introduction to the field of antibiotics and antibiotic resistance for students or interdisciplinary collaborators.