RADseq for Antarctic Usnea species

RADseq libraries for Usneaantarctica and U.aurantiacoatra were prepared as described previously (Grewe et al. 2017 (link)). In short, for the RADseq library production, DNA isolations were pooled with sequence adapters (Rubin and Moreau 2016 (link)), subsequently digested with the restriction enzyme ApeKI (New England Biolabs, Ipswich, MA, USA) and ligated using T4 ligase (New England Biolabs). Up to 48 samples with compatible barcodes were pooled and selected for fragments of sizes between 300 and 500 bp using the BluePippin DNA size selection system (Sage Science, Beverly, MA, USA). The pooled libraries were amplified using the REDTaq ReadyMix (Sigma-Aldrich, St. Louis, MO, USA) prior to sequencing on an Illumina MiSeq using the MiSeq Reagent Kit v3 for 150 cycles (Illumina, San Diego, CA, USA) to produce single-end sequences with a length of 150 bp.