CHALLENGES ASSOCIATED WITH BARCODING METHOD

Abstract

The DNA barcoding project provides a very quick and easy way to identify different types of organisms based on their DNA. The main objective of this study is to obtain the sequences of different species of fungi, plants and animals to map the biodiversity of Slovakia. This project also represents a new use for collections stored in natural history museums around the world. This paper shows in particular a laboratory view of the DNA barcoding project, such as the practical application of the methodology for different groups of organisms, the possibilities of using different genes and emphasizes the importance of professional taxonomists. In the museum, we deal with a wide range of samples and we encounter various difficulties. For example, some genera of fungi contain unreadable sections in the middle of the sequence, in insect groups it is a problem with samples killed with ethyl acetate or too old samples, where the DNA is mostly degraded. For the amplification of bird samples, we have found no consistent method for the whole group and we work with several different primer sets and conditions to be able to amplify most of the samples. The identification of various plant species according to their barcodes has also proven quite difficult, as plants are a very specific and fast-evolving group of organisms, and their distinction according to the short barcode regions is nearly impossible. Therefore, we would like to emphasize the necessity of cooperation with specialized taxonomists. Our data are continuously uploaded to the international BOLD database, where there are already more than 500 different species of a wide range of groups of fungi, plants and animals from Slovakia.