Background: Neck pain is one of the most common causes of consultation in general practice. The DEGAM (“Deutsche Gesellschaft für Allgemeinmedizin”) S3 guideline “Nonspecific neck pain” was developed to support evidence-based management of diagnosis and treatment. The acceptance, practicability and implementability of the guideline and the associated patient information were evaluated as part of the practice test. The evaluation of the guideline by means of a practice test is a special feature of the DEGAM guideline development. Materials and methods: Mixed methods approach consisting of questionnaire surveys (general practitioners [GPs] and patients [PAT]) and focus group discussions (GPs) was used. The questionnaires were developed independently and were thematically based on the DEGAM guideline authors’ manual. The GPs tested the long and short versions of the guideline for 7 weeks and documented patient contacts with nonspecific neck pain. Furthermore, the physicians handed out the patient information and an evaluation form to the PATs. Questionnaires were analyzed descriptively. The two focus group discussions were semi-structured and the evaluation was based on Mayring’s content analysis using a thematic summary. Results: A total of 11 GPs documented 86 patient contacts and 10 GPs evaluated the guideline. In all, 59 PAT participated in the patient survey. The GPs judged the guideline to be helpful in 65/86 cases and 53/59 PAT assessed the patient information as at least good. Challenges in applying the guideline arose from patient-specific therapy expectations (e.g., physiotherapy), and deviations from the guideline were justified, among other things, by experience-based knowledge. Therapeutic formats (including acupuncture and taping) used in the GP practice were either insufficiently explained or not explained at all in the guideline. Conclusion: The practice test showed that the guideline provides useful support for primary care. Expanding the guideline to include more detailed explanations and visual materials could further increase acceptance and adherence.