Location: Floral and Nursery Plants ResearchTitle: Optimizing efficient PCR-amplifiable DNA extraction from herbarium specimens
Submitted to: Applications in Plant Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/11/2023
Publication Date: 5/22/2023
Citation: Gouker, F.E., Guo, Y.H., Svoboda, H.T., Pooler, M.R. 2023. Optimizing efficient PCR-amplifiable DNA extraction from herbarium specimens. Applications in Plant Sciences. https://doi.org/10.1002/aps3.11521.
Interpretive Summary: Herbarium specimens are becoming increasingly recognized as a vital resource to help answer research questions related to understanding taxonomic identity, evolutionary relationships, genetic diversity, and many other aspects across the biological sciences. Preserved collections also afford the opportunity to study taxa from a wide geographic distribution, as well as those that are extinct or even undescribed. However, DNA extraction from herbarium samples is often difficult due to degradation of the specimen, plant chemistry, morphological characteristics, or method of collection and preservation. Although several methods have been developed to address these challenges, many protocols are often complicated and/or rely on expensive kits. ARS scientists developed a fast, easy, cost-effective method that utilizes common laboratory chemicals and supplies and has been optimized for extraction of DNA from herbarium specimens of varying age and condition.
Technical Abstract: Our lab developed a rapid and simple acetone-based DNA extraction protocol, and we optimized the protocol for use on herbarium tissue. Leaf tissue of 30 diverse plant species was obtained from the U.S. National Arboretum Herbarium, and DNA was extracted using this new method, which included a combination of SDS, 10X TE buffer, PVPP. DNA yields were greater than 110 ng/mg tissue for all samples and PCR amplification thresholds (Ct values) using several universal plant primers showed DNA amplification for most of the specimens. This protocol, which is simple, fast, and uses standard laboratory-grade chemicals, yields DNA from herbarium specimens that is comparable in quality to that from commercially available kits, and is of sufficient quality and quantity for downstream applications.