Three steps towards comparability and standardization among molecular methods for characterizing insect communities. 2024

Ela Iwaszkiewicz-Eggebrecht, and Vera Zizka, and Christina Lynggaard
Bioinformatics and Genetics Department, Swedish Museum of Natural History, PO Box 50007, Stockholm, 104 05, Sweden.

Molecular methods are currently some of the best-suited technologies for implementation in insect monitoring. However, the field is developing rapidly and lacks agreement on methodology or community standards. To apply DNA-based methods in large-scale monitoring, and to gain insight across commensurate data, we need easy-to-implement standards that improve data comparability. Here, we provide three recommendations for how to improve and harmonize efforts in biodiversity assessment and monitoring via metabarcoding: (i) we should adopt the use of synthetic spike-ins, which will act as positive controls and internal standards; (ii) we should consider using several markers through a multiplex polymerase chain reaction (PCR) approach; and (iii) we should commit to the publication and transparency of all protocol-associated metadata in a standardized fashion. For (i), we provide a ready-to-use recipe for synthetic cytochrome c oxidase spike-ins, which enable between-sample comparisons. For (ii), we propose two gene regions for the implementation of multiplex PCR approaches, thereby achieving a more comprehensive community description. For (iii), we offer guidelines for transparent and unified reporting of field, wet-laboratory and dry-laboratory procedures, as a key to making comparisons between studies. Together, we feel that these three advances will result in joint quality and calibration standards rather than the current laboratory-specific proof of concepts. This article is part of the theme issue 'Towards a toolkit for global insect biodiversity monitoring'.

UI MeSH Term Description Entries
D007313 Insecta Members of the phylum ARTHROPODA composed or organisms characterized by division into three parts: head, thorax, and abdomen. They are the dominant group of animals on earth with several hundred thousand different kinds. Three orders, HEMIPTERA; DIPTERA; and SIPHONAPTERA; are of medical interest in that they cause disease in humans and animals. (From Borror et al., An Introduction to the Study of Insects, 4th ed, p1). Insects,Insect
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D044822 Biodiversity The variety of all native living organisms and their various forms and interrelationships. Biological Diversity,Diversity, Biological
D058893 DNA Barcoding, Taxonomic Techniques for standardizing and expediting taxonomic identification or classification of organisms that are based on deciphering the sequence of one or a few regions of DNA known as the "DNA barcode". DNA Barcode, Taxonomic,Phylogenetic DNA Barcode,Phylogenetic DNA Barcoding,Taxonomic DNA Barcode,Taxonomic DNA Barcodes,Taxonomic DNA Barcoding,Barcode, Phylogenetic DNA,Barcode, Taxonomic DNA,Barcodes, Phylogenetic DNA,Barcodes, Taxonomic DNA,Barcoding, Phylogenetic DNA,Barcoding, Taxonomic DNA,Barcodings, Phylogenetic DNA,Barcodings, Taxonomic DNA,DNA Barcode, Phylogenetic,DNA Barcodes, Phylogenetic,DNA Barcodes, Taxonomic,DNA Barcoding, Phylogenetic,DNA Barcodings, Phylogenetic,DNA Barcodings, Taxonomic,Phylogenetic DNA Barcodes,Phylogenetic DNA Barcodings,Taxonomic DNA Barcodings
D060885 Multiplex Polymerase Chain Reaction Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction. Multiplex Ligation-Dependent Probe Amplification,Multiplex PCR,Triplex PCR,Triplex Polymerase Chain Reaction,Multiplex Ligation Dependent Probe Amplification,PCR, Multiplex,PCR, Triplex

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