Thornicroft’s giraffe, Giraffa camelopardalis thornicrofti, is a geographically isolated subspecies of giraffe found only in north-east Zambia. The population only occurs in Zambia’s South Luangwa Valley, an area which interestingly places it between the current distribution of Masai (G. c. tippelskirchi) giraffe to the north, and the Angolan (G. c. angolensis) and South African (G. c. giraffa) giraffe in the south-west and south, respectively. Speciﬁc studies have been undertaken on the ecology of this subspecies, but their population genetics remains unknown. We studied 34 individuals from the South Luangwa National Park and adjacent Lupande Game Management Area and seven individuals from northern Botswana. The complete cytochrome b and control region sequences of the mitochondrial genome were sequenced and analysed together with database data by maximum likelihood tree reconstruction and maximum parsimony network analyses. The giraffe from Zambia’s South Luangwa Valley are most closely related to the subspecies G. c. tippelskirchi and part of their radiation. However, they form a unique population that would beneﬁt from increased research and conservation management.