AbstractNematode‐trapping fungi (NTF) and nematodes are common and sympatric in nature. The molecular basis that underlies this interkingdom predator–prey interaction remains largely uncharacterized. Both NTF and nematodes can be easily isolated from soil samples. NTF do not form traps in nutrient‐rich environments, yet trap morphogenesis can be observed under nutrient‐poor conditions and upon simultaneous sensing of the nematode cues. Here, we present protocols for laboratory maintenance and culturing of the model NTF Arthrobotrys oligospora. © 2021 Wiley Periodicals LLC.This article was corrected on 19 July 2022. See the end of the full text for details.Basic Protocol 1: Growth of nematode‐trapping fungi on solid mediumBasic Protocol 2: Growth of nematode‐trapping fungi in liquid mediumBasic Protocol 3: Collection of conidia from solid mediumSupport Protocol 1: Preparation of Miracloth filter funnelBasic Protocol 4: Induction of trap morphogenesisAlternate Protocol: Quantitative measurement of trap inductionSupport Protocol 2: Preparation of synchronized C. elegans L4Basic Protocol 5: Establishing C. elegans survival rate upon exposure to A. oligosporaBasic Protocol 6: Storage of nematode‐trapping fungi strains