The literature states that transitional vertebrae at any junction are characterized by features retained from two adjacent regions in the vertebral column. Currently, there is no published literature available that describes the prevalence or morphology of thoracolumbar transitional vertebrae (TLTV). The aim of this study was to identify the qualitative characteristics of transitional vertebrae at the thoracolumbar junction and establish a technique to differentiate the various subtypes that may be found. A selection of vertebral columns from skeletal remains (n = 35) were evaluated in this study. Vertebrae were taken based on features that are atypical for vertebrae in each relative region. The transitional vertebrae were qualitatively identified based on overlapping thoracic and lumbar features of vertebrae at the thoracolumbar junction. The following general overlapping characteristics were observed: aplasia or hypoplasia of the transverse process, irregular orientation on the superior articular process and atypical mammillary bodies. The results show that the most frequent location of the transitional vertebrae was in the thoracic region (f = 23). The second most frequent location was in the lumbar region (f = 10). In two specimens of the selection (f = 2), an additional 13th thoracic vertebra was present which functioned as a transitional vertebra. This study concluded that one can accurately identify the characteristics of transitional vertebrae at the thoracolumbar junction. In addition, the various subtypes can be differentiated according to the region in the vertebral column the vertebra is located in and the relative number of vertebral segments in the adjacent regions of the vertebral column. This provides a qualitative tool for researchers to differentiate the transitional vertebrae from distinctly different typical thoracic or lumbar vertebrae at the thoracolumbar junction.