Note on using nuclear 28S rDNA for sequencing ancient and strongly degraded insect DNA

摘要

A protocol using insect specimens or parts thereof allows for sequencing of sections of nuclear 28S rDNA. In the present note it is demonstrated that this protocol can readily be applied to strongly degraded DNA (ancient, fixed or contaminated). Primers that are specifically designed to discriminate against humanDNAbut also other non-arthropod species are tested on a range of species covering all insect groups (59 insect species from all 33 orders). Additionally, the samples represent a selection of various, mostly DNA-degrading, preservation methods, including the most common fixatives used for morphological investigations and for long-term storage in collections. Successful amplification was possible for all tested samples including ca. 200 year-old dried museum specimens as well as for over 4000 year-old fossil insects embedded in copal. When the NCBI database contained information on the tested species an unambiguous taxonomic discrimination was possible. This approach is based on a standardized protocol that guarantees easy application. This note presents primer pairs for 28S rDNA that can be a useful tool for ancient DNA (aDNA) research.