DNA extraction from frozen field-collected and dehydrated herbarium fungal basidiomata: performance of SDS and CTAB-based methods

Abstract

Extracted DNA from basidiomata of frozen field-collected and herbarium specimens collected in different periods and maintained in a variety of ways was evaluated according to general condition, color, spectral absorbance ratio, final concentration and PCR amplification, using two distinct and most common extraction methods: SDS (Sodium Doecyl Sulphate) and CTAB (Cetyl Trimetyl Ammonium Bromide). Most of the frozen fieldcollected basidiomata presented a very good to good DNA condition and no degradation. All the herbarium specimens exhibited poor DNA condition with low molecular weight bands. Practically all the DNA solutions of the samples were colored, and spectral absorbance ratios were considerably variable, with lower values observed in darker ones. All the samples were successfully PCR-amplified. DNA extraction of Herbarium specimens and also specimens with high polysaccharide content was only achievable using the CTAB method. For an all-purpose DNA extraction from field-collected and dehydrated coriaceous and woody basidiomata bearing a high content of secondary metabolites and/or polysaccharides, the CTAB-based method is definitely best the choice of method.

Keywords

• PCR methods
• basidiome
• Basidiomycetes