Trehalose improves PCR amplification of vertebrate nuclear DNA from historical allozymes
Historical tissue collections represent potential resources for temporal genetic studies in evolutionary and conservation biology. Unfortunately, DNA from historical samples stored without modern genetic analyses in mind, such as frozen allozyme homogenates, are often degraded and contaminated with PCR inhibitors. Here, we report the successful use of trehalose for improving PCR amplification of degraded, vertebrate nuclear DNA extracted from cryopreserved allozymes. We amplified and sequenced two nuclear genes (MLC2a and RPL12) from allozymes of the Shasta salamander, Hydromantes shastae. Our results demonstrate the potential of trehalose as a tool for utilizing historical allozyme collections in modern genetic studies.
KeywordsConservation Degraded DNA PCR enhancer Museum samples Salamander
We thank David Wake, Carol Spencer, and the Museum of Vertebrate Zoology (MVZ) for allozyme and tissue samples. We thank the MVZ’s Evolutionary Genetics Lab and Lydia Smith for access to sequencing facilities. We also thank an anonymous reviewer for providing thoughtful feedback on this manuscript. This work was supported by a grant from the Hellman Fellows Fund, the California Agricultural Experiment Station, and the USDA National Institute of Food and Agriculture, Hatch project 1007819.
- R Development Core Team (2017) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. http://www.R-project.org/