We used ancient DNA sequencing to assign mitochondrial DNA (mtDNA) haplogroups to 29 individuals from a 19th century Primitive Methodist burial ground in Darwen, Lancashire, UK. The burial ground contained at least 142 skeletons distributed among 74 grave plots, just under half of which were multiple stacked interments. For five of the eight plots from which we studied multiple burials, the mtDNA haplogroups were consistent with maternal and/or sibling relationships among the co-interred individuals, supporting the hypothesis that individual burial plots contained members of the same family. The majority of the haplogroups that were identified have distributions that include northwest Europe and hence are not unexpected in a 19th century burial group from northern England. Exceptions were a female adult and two male subadults, from a single burial plot, who possessed haplogroup U6a1a, which is associated with northwest African populations and is less commonly found in Iberia and southern Italy, and a subadult of unknown sex who was tentatively assigned haplogroup A12a, which is primarily found in Native American populations. The results therefore provide an insight into the mobility and rich genetic makeup of the poor working class people of industrial Britain in the 19th century.