Cultivation and Genomic DNA Extraction of Klebsiella pneumoniae

Abstract Klebsiella pneumoniae is a Gram‐negative, rod‐shaped bacterium of medical significance. It typically exists as part of the normal flora of the human intestine but can cause severe infections in the healthcare setting due to its rapid acquisition of antibiotic resistance. Cultivating and extracting genomic DNA from this bacterium is crucial for downstream characterization and comparative analyses. To provide a standardized approach for growing K. pneumoniae in the laboratory setting, this collection of protocols provides step‐by‐step procedures for routine culturing, generating growth curves, storing bacteria, and extracting genomic DNA. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1 : Reviving K. pneumoniae from frozen stocks Basic Protocol 2 : Cultivating K. pneumoniae in rich growth medium Alternate Protocol : Cultivating in minimal liquid growth medium Basic Protocol 3 : Enumerating K. pneumoniae colony forming units Basic Protocol 4 : Growth curves Basic Protocol 5 : Genomic DNA extraction Basic Protocol 6 : Characterizing K. pneumoniae strains based on genomic sequence Basic Protocol 7 : Storage of K. pneumoniae frozen stocks in glycerol Basic Protocol 8 : Storage of K. pneumoniae in agar stabs