Amplification and Sequencing

Amplification and sequencing of DNA from fungal herbarium specimens.

Bruns, T D; Fogel, R; Taylor, J W
Department of Plant Pathology, University of California Berkeley 94720.
Mycologia. 1990 82(2):175-184.

Abstract
We have utilized the polymerase chain reaction to amplify DNA from 35 collections of dried fungi obtained from four different herbaria. Sixteen of these samples were subsequently sequenced. The collections had been dried and stored under a variety of conditions for 1-50 years. DNA was extracted from 5-30 mg of ground tissue and a specific fragment of the mitochondrial large subunit ribosomal RNA gene was amplified. Sequences obtained from DNA cloned from a culture and those from DNA amplified from the corresponding dried voucher collection were found to be identical. Controls lacking DNA failed to produce amplified products. This application of the polymerase chain reaction increases the value of fungal herbarium specimens by enabling their use in molecular systematic and population genetic studies, and it creates new curatorial demands.