Quick n’ Cheap – a simplified workflow to barcode plasmodial slime molds (Myxomycetes) Martin Schnittler 1* , Nikki H.A. Dagamac 1,2 , Dmitry Leontyev 3 , Oleg Shchepin 1,4 , Yuri K. Novozhilov 4 , Anja Klahr 1 1 Institute of Botany and Landscape Ecology, University Greifswald, Soldmannstr. 15, 17489, Greifswald, Germany 2 Department of Biological Sciences and Research Center for the Natural and Applied Sciences, University of Santo Tomas, España 1008 Manila, Philippines 3 Department of Botany, H.S. Skovoroda Kharkiv National Pedagogical University, Valentynivska 2, Kharkiv 61168 Ukraine 4 V.L. Komarov Botanical Institute of the Russian Academy of Sciences, Prof. Popov St. 2, 197376 St. Petersburg, Russia * Corresponding author: martin.schnittler@uni-greifswald.de Keywords: DNA barcoding, DNA extraction, elonga- tion factor 1 alpha, direct PCR, spore collection, small ribosomal subunit, spore. Article info: Received: 26 November 2020 Accepted: 13 December 2020 Published online: 31 December 2020 Corresponding Editor: Riikka Linnakoski Abstract We present a workfow for efcient barcoding of myxomycete fructifcations, which (i) requires less than 1000 spores, (ii) allows to collect spores with only a needle, (iii) works without any commercial kits, and (iv) is optimized for the use of 96-well PCR plates throughout the process. Specimens of 291 dark-spored nivicolous myxomycetes and 121 bright-spored members of the Trichiaceae were sequenced for the barcode marker 18S rDNA (SSU) with a low rate of failure and no detectable cross-contamination. Crude DNA extracts can be stored for further analyses: the elongation factor 1 alpha gene (EF1A), a single-copy marker, was successfully amplifed after four weeks of storage. As such our procedure will allow a time- and cost-ef- fcient barcoding of large series of specimens. doi:10.29203/ka.2020.504 Karstenia, Volume 58 (2020), Issue 2, pages 385–392 www.karstenia.f METHODS AND PROTOCOLS