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WGS of Ustilago bromivora x Ustilago hordei fungal hybrids
Extracted fungal gDNA was adjusted to 2.5 ng/l after quantification using picogreen dye (Thermo Fisher Scientific) to exclusively measure double-stranded DNA. We followed a modification of the Illumina library preparation protocol prepared by Sergey Kryazhimskiy (V. 2.1 (2013-06-06)) which was later published, after further modifications, by Baym et. al. Barcoding was done using an Illumina Nextera kit (Illumina) for labelling 24 samples. We performed library amplification using either Veraseq polymerase (Biozyme) or Phusion polymerase (Thermo Fisher Scientific) and size selection with AMPure XP beads (Beckman Coulter). Fragment profiles of the libraries were determined with a Fragment Analyzer (Advanced Analytical) and libraries were pooled to equal concentrations, either based on the concentration determined by the Fragment Analyzer or a second round of quantification with picogreen fluorescent spectrometry. The libraries were sequenced using Illumina HiSeqV4 PE125 on a HiSeq2500 at the VBCF NGS Unit (www.vbcf.ac.at).
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SUB4878542
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