This work aims at fractography of polyamide 66 nanofiber yarns. The yarns are produced with three twist levels via electrospinning. In order to study the fracture modes of nanofiber yarns, fatigue, and static tensile tests including monotonic, low cycle fatigue, and postcyclic monotonic tensile tests are performed. It is observed that the catastrophic failure of yarns is associated with axial splitting in the three categories. The nanofibers within the yarn structure show a ductile fracture and buckle after tensile stress release. In comparison of postcyclic monotonic tensile tests with other categories, nanofibers show severe plastic buckling in response to release of the same applied force. Fractography studies reveal that twisting causes construction of a layered structure in the yarns which is similar to the ideal yarn structure as well. Applying cyclic loading causes the separation of these structural layers which is more considerable under higher number of cycles. (c) 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 41925.