Fecal microbiota transplant (FMT) is a therapeutic modality that targets intestinal microorganisms by transplanting fecal microorganisms from the donor into the recipient's gastrointestinal tract in order to reconstitute the recipient's intestinal flora. However, the mechanism of action and adverse effects of FMT in response to different diseases have not yet been clarified, and thus it cannot be widely applied in the clinic, and still needs to rely on in-depth preclinical studies. The highly inconsistent or incomplete experimental details in current reports, coupled with the lack of authoritative standards and recommendations, not only seriously affect the interpretation of their findings and the replication of experimental procedures, but also hinder the clinical translation of the results. We review and discuss the key steps of recipient selection and graft sample collection, storage, graft material preparation, and grafting route, with the aim of improving the utilization of experimental animals, consumables, and labor, and providing methodological recommendations and references to achieve replicability and standardization of FMT preclinical studies.