, 2003). This technique has the advantage of being independent of the user’s taxonomic expertise and makes it possible to assign species names to specimens or samples that are challenging (or impossible) to identify any other way. Importantly, this applies not only individual organisms (or tissues from those organisms, like a fin clip from Veliparib mw a fish or leg from a crab), but also to environmental or ‘bulk’ samples, from which the target gene/barcode

can be sequenced. The approach consisting in sequencing a DNA fragment from a whole environmental sample is sometimes called metagenetics or metabarcoding (for example, see: Taberlet et al., 2012). The essential prerequisite for DNA barcoding (and metabarcoding) is the creation of a reference database consisting of a library of species names linked to the DNA barcodes. Building the reference library requires an expert taxonomist to name a representative specimen for each species (usually deposited in a natural history museum or

herbarium) and to sequence the specimen for the appropriate barcode gene (or genes) designated by the international Consortium for the Barcode of Life (CBOL). The reference library (usually created from adult life stages) serves as a tool for robust and reproducible species identification for assigning biological material (any sample with DNA) to species so long as the DNA barcode can be sequenced from the sample and is present MK-1775 mw in the reference library. The BOLD platform (http://www.barcodinglife.com), which is one of the largest existing DNA barcode libraries, contains over two million sequences (as of February 2013), of which almost 130,000 are formally described animals, over 42,000 are formally described plants and about 2500 are formally described fungi and protists 17-DMAG (Alvespimycin) HCl (Hajibabaei, 2007). DNA barcoding techniques have the potential to contribute to a large number of MSFD indicators (Table 3) and other legislation worldwide, wherever

species identification is required, such as indicators of biological diversity, non-indigenous species, and food webs. DNA barcoding and metabarcoding have a high priority for marine monitoring and assessment, and more pilot studies and cost-benefit analyzes are needed to test the general applicability of this method. In 2006, the cost of DNA barcoding was estimated at about $5 per sample (Cameron et al., 2006), including: DNA extraction, US$1.90; PCR, US$0.37; PCR purification, US$0.28; and Sanger sequencing, US$2.36, plus minor laboratory supplies such as buffers, gels, etc. Note that this does not include the collection or transport of the specimen or sample and it assumes that the species is already present in a reference library.