Spiders have strong effect on the density of insect populations and have been shown to limit insect pests in the agricultural environment. On reviewing literature that focused on the Egyptian spider's fauna, it could be concluded that the correct identification of many spider species is considered problematic due to complex variations in morphology and lack of systematic studies. Thus, the present study was aimed to design a key to facilitate identification of true spiders using their morphological characteristics and make a catalogue for different families and species inhabiting Assiut governorate; the central part of Upper Egypt (27º14' N and 31º11' E) and lies within the arid belt of North Africa.  Samples of spiders were collected monthly during a period of one year (from December 2015 till November 2016) from six different sites covering Assiut governorate using hand picking method. Survey results revealed the occurrence of 3457 specimens belonging to 22 families that included 42 genera and 47 species. Out of these species, 40 species are considered as new locality records at Assiut Governorate.