The medical laboratory science (MLS) profession is in need for published molecular diagnostics competency-based standards and curriculum. To assess their expectations of new MLS graduates, professionals performing and supervising performance of clinical molecular assays were surveyed to rate the importance of relevant cognitive and psychomotor learning objectives. A modified, asynchronous, iterative online Delphi process was utilized for assessment of consensus on the importance of the objectives. The survey was delivered through online REDCap application. Program directors of 221 MLS programs accredited by the National Accrediting Agency for Clinical Laboratory Science (NAACLS) were asked to forward the first Delphi survey to target participants at their affiliated clinical sites. Ninety-four experts submitted complete surveys, including 88 who provided email addresses, indicating agreement to participate in future Delphi rounds. Most of the participants were certified by ASCP or NCA (81.9%), had over 10 years of laboratory experience (76.6%), and worked in a hospital setting (43.6%). The reliability of the surveys, assessed using Cronbach’s alpha, was 0.96 and 0.97. In the second survey, the objectives assigned low importance by the majority were removed; and others, assigned high importance were expanded. Respondents were given the opportunity to confirm or change their opinion on the objectives after reviewing quantitative results and narrative comments collected in the preceding survey. Upon completion of the Delphi process, 25 essential items were identified as necessary for inclusion in the entry-level MLS curriculum. These concepts and objectives focused on basic molecular biology principles and general molecular laboratory operations, including practical knowledge of techniques designed to maintain specimen integrity and intense theoretical background of the polymerase chain reaction, as well as comprehension of the principles of laboratory assays designed for pathogens most commonly tested for using molecular methods. In this study, the investigator also provided information on the preferred number of contact hours devoted to each group of the identified essential items. The goal of creating the list of essential concepts and objectives was to share it with MLS educators, the NAACLS and the provider of MLS certification exam, the American Society for Clinical Pathology Board of Certification (ASCP-BOC), to contribute to the existing exam content guidelines.