Modified method of DNA extraction from herbarium specimens

Abstract

The DNA isolation CTAB-method modified for seed plants herbarium material is proposed. Three modifications were tested on 29 samples from KW and KWU herbaria. Visual evaluation of optimal method is done for DNA containing pellet, PCR products (nuclear ITS and plastid rbcL gene sequence), amplicon sequencing with further comparing results obtained using those deposited in GenBank. The optimal variant of DNA extraction from herbarium material for different taxonomical groups of plants is proposed.

Resolving the position of Astragalus borysthenicus Klokov within the Astragalus L. species

“…Total genomic DNA was extracted from herbarium material using a modification (Tarieiev et al, 2011) of the method developed by Doyle and Doyle (1990). PCR reactions of ITS1-5.8S-ITS2 regions were performed as described by White (1990) using ITS1 and ITS4 universal primers.…”

The Promise of Molecular and Genomic Techniques for Biodiversity Research and DNA Barcoding of the Arabian Peninsula Flora

“…It is also highlighted that the process of extracting DNA from herbarium specimens is often fraught with difficulty related to such variables as plant chemistry, specimen drying methods, and the chemical treatment of specimens (Drábková, 2014). Thus far, many methods have been developed for the extraction of DNA from herbarium specimens, with the most frequently used being either the traditional CTAB protocol (Doyle and Dickson, 1987), sometimes with modifications (Allen et al, 2006; Cota-Sánchez et al, 2006; Tarieiev et al, 2011) or DNA extraction kits such as the DNeasy Plant Mini Kit (Qiagen) (Drábková, 2014). Särkinen et al (2012) outlined the major challenges of molecular studies using herbarium DNA and emphasized that despite the large number of specimens housed in herbaria worldwide, currently only a small fraction is being used for DNA-based research, mainly due to the poor success and difficulties in obtaining amplifiable DNA.…”

“…In recent years, the number of phylogenetic studies using herbarium specimens has been gaining momentum. Researchers have been using herbarium specimens in molecular phylogeny studies and consider them as an interesting potential source of material for DNA barcoding and the construction of a barcode library for flora (Cozzolino et al, 2007; Erkens et al, 2008; Tarieiev et al, 2011; de Vere et al, 2012; Särkinen et al, 2012; Hebert et al, 2013; Xu et al, 2015; Bakker, 2017). In such studies, collections of herbaria are used when species are not easily collected in the field, since it is generally difficult to obtain living material of certain rare species (Délye et al, 2013).…”

“…Additionally, the quality of DNA obtained from herbarium specimens should ideally be consistent with freshly collected samples for barcoding studies. Some researchers have successfully amplified DNA from herbarium specimens by modifying methods of DNA extraction (Cota-Sánchez et al, 2006; Andreasen et al, 2009; Staats et al, 2011; Tarieiev et al, 2011; Gutaker et al, 2017). However, some researchers have highlighted that it is challenging to find a suitable genomic region for DNA barcoding in herbarium samples in a wide range of taxa (Lahaye et al, 2008a,b; Poczai et al, 2009; Drábková, 2014).…”

Morphological, Karyological and Molecular Characteristics of Festuca arietina Klok. – a Neglected Psammophilous Species of the Festuca valesiaca agg. from Eastern Europe

“…Leaf fragments of herbarium specimens were used for molecular analysis. Total DNA extraction was performed using a CTAB-method (Doyle, Doyle, 1987) modified for herbarium specimens (Tarieiev et al, 2011 manually using BioEdit software (Hall, 1999). The obtained sequences were deposited in NCBI (accession numbers KP796238 (Ukraine, locus classicus) and KP796239 (Belarus)).…”