Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 6 July 2021 doi:10.20944/preprints202107.0151.v1 Article Universal Set of Primers for DNA Identification of Alien Fish Species in Central Russia (Volga-Kama region) Dmitry P. Karabanov 1,*, Eugenia I. Bekker 2, Dmitry D. Pavlov 1, Elena A. Borovikova 1, Yulia V. Kodukhova 1, and Alexey A. Kotov 2,* 1 I. D. Papanin Institute for Biology of Inland Waters of Russian Academy of Sciences, Borok Yaroslavl Area, 152742, Russia;
[email protected] 2 A. N. Severtsov Institute of Ecology and Evolution of Russian Academy of Sciences, Leninsky Prospect 33, Moscow 119071, Russia;
[email protected] * Correspondence:
[email protected] (DPK),
[email protected] (AAK) Abstract: Reliable species identification is critical for detection and monitoring of biological inva- sions. In this study, we propose four sets of primers for efficient amplification of several loci, in- cluding the mitochondrial cytochrome oxidase-c (COI) subunit I gene which is a basis for DNA barcoding. This set of primers gives a shorter product which can be used in high-throughput se- quencing systems for metabarcoding purposes. Another mitochondrial locus encoding the large ribosomal subunit (16S) may be useful to study the population structure and as an additional source of information in the metabarcoding of communities. We propose to use a set of primers for the nuclear locus of the small ribosomal subunit (18S) as a positive control and to verify the results of the barcoding. Our proposed sets of primers demonstrate a high amplification efficiency and a high specificity both for freshwater alien and indigenous fishes.