Quick n' Cheap - A simplified workflow to barcode plasmodial slime molds (Myxomycetes)

Abstract

We present a workflow for efficient barcoding of myxomycete fructifications, 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 amplified after four weeks of storage. As such our procedure will allow a time- and cost-efficient barcoding of large series of specimens.