Background: The Franciscan Friary in Montella near Avellino in Southern Italy is of special interest because according to historical sources it was founded by St. Francis himself in AD 1221-1222. Human remains of several hundred individuals interred in the cloister walk have been unearthed during two excavation campaigns conducted in 2007-2008 and 2010. The environs of the friary have remained rural since the foundation preventing much modern contamination. The state of preservation of the skeletons is fair to good making a suite of analyses worthwhile. Results: The skeletons have been examined anthropologically and tissue samples have been subjected to radiocarbon dating, stable isotope measurements and trace element analyses by Inductively Coupled Plasma Mass Spectrometry and Cold Vapour Atomic Absorption Spectrometry. Conclusions: The radiocarbon dates are consistent with the historical sources and show that the cemetery in the cloister walk has been in uninterrupted use from the foundation of the friary in AD 1221-1222 and until the cemetery went out of use in AD 1524. The anthropological investigations show that the individuals interred at the friary would have been shorter than other Italians from the same time, and it seems that tuberculosis was more prevalent than leprosy. Isotopic measurements show a mixed agricultural and pastoral diet and none of the individuals were consuming marine protein. Based on the trace element analysis it seems that the people resided mainly at two distinct geographical areas, one of which was Montella. One individual stands out from the rest, because he was born and raised at some third geographical location distinct from Montella and because he sports the second oldest radiocarbon date of AD 1050-1249 (two sigma calibrated range). This date is consistent with the first generation of the founders of the friary-perhaps one of St. Francis' fellow travellers from Assisi.