However, accurate and consistent taxon identification has proved difficult to achieve using traditional morphological approaches. This is particularly true for the largescale application of macroinvertebrate sampling in river biomonitoring, where larval stages are often difficult or impossible to identify below the level of taxonomic family. This issue has caused difficulties in implementing large-scale biomonitoring programs, particularly in relatively less-populated countries such as Canada, where remoteness poses a significant logistic challenge for sample collection, coupled with poor knowledge of the local fauna. Sanger��s invention of DNA sequencing revolutionized all branches of the biological sciences. In biosystematics, DNA sequence information provides vast amounts of reproducible and robust genetic data that can be informative at nearly any level of taxonomic hierarchy: from individuals in populations, to species, to the deepest branches of the Tree of Life. DNA sequence-based analyses have provided evolutionary biologists and ecologists the opportunity to address questions they could not answer using other types of data. In recent years��particularly with the introduction of the concept of DNA barcoding in 2003 ��efforts have been directed towards building a FK-3311 standard sequence library for all eukaryotes by focusing DNA sequencing efforts on small, speciesspecific portions of the genome called DNA barcodes. The primary utility of DNA barcoding is to identify unknown specimens at the species-level by comparing the query sequence to a DNA barcode reference library built based on known species. In addition, patterns of sequence variation can be used to flag new and cryptic species. By sampling more genes or individuals, DNA barcode projects can shift to population-level analysis or deep phylogenetic questions. In the past seven years, over 1.1 M individuals from about 95,000 species have been added to the DNA barcode library. This number is not significant in the context of the 1.9 M known and 10�C100 M estimated unknown species. However, this progress is significant because DNA barcoding in the past seven years has chiefly been geared towards proof-of-concept projects to enhance application through the development of improved protocols. Major hurdles in the high-throughput analysis of DNA barcodes have been resolved and single analytical facilities can now Soyasaponin-Bb process several hundred thousand samples per year. Global projects such as the International Barcode of Life project and other concerted efforts to barcode taxonomic groups or regional biota will rapidly increase the sequence coverage in DNA barcode libraries. Although Sanger-based DNA sequencing has proved robust for building large sequence libraries such as DNA barcode reference libraries, it is not a feasible approach for tackling bulk environmental samples because these samples can contain thousands of individuals from hundreds of species ranging from bacteria to higher eukaryotes. Separating these individuals and then using single-specimen Sanger sequencing has historically been challenging and for some material is beyond the scope of traditional technologies. Although cloning followed by sequencing a library of cloned fragments partially addresses this problem, this method has its own limitations and can introduce biases. Consequently, biomonitoring programs and other large-scale biodiversity analyses in ecological and environmental studies cannot be performed routinely on a large-scale using a single-specimen Sanger sequencing workflow.