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The Chequered Skipper butterfly (Carterocephalus palaemon) is extinct in England but extant in Scotland and Continental Europe. The possibility of re-introducing the species is under active consideration by conservation bodies, but ecological differences between Scottish and English populations raise the question of which populations should donate individuals, Continental European, or Scottish? We used mitochondrial DNA sequences (CO I, CO II and Cyt b) from potential donor populations to test the hypothesis that ecological differences could have arisen as a result of differing routes of post-glacial colonisation from separate refugia and subsequent isolation of UK populations. Shared haplotypes between populations in Belgium, Norway, Scotland and England provides no evidence to support the hypothesis that populations in Scotland result from an alternative post-glacial colonisation route. As the genetic evidence remains equivocal we suggest that choice of donor stock for a re-introduction to England should be made primarily on ecological grounds. [ABSTRACT FROM AUTHOR]