A plant visibly infected with both Hyaloperonospora arabidopsidis and Albugo sp. was collected in Fall 2014 from the village of Gniebel (48° 34′ 34.10″ North Lat., 9° 10′ 55.42″ East Long.) using sterile tweezers and scissors, placed in a sterile 15 mL tube, and brought back to the lab on ice where it was frozen at −80 °C until further processing. The frozen plant was ground in the presence of liquid nitrogen using a mortar and pestle that was lined with four layers of autoclaved aluminum foil. Approximately 250 g of the resulting powder was used for DNA extraction, using a custom protocol we previously described [22 (link)]. Briefly, the sample was subjected to bead-beating in the presence of 1.5% sodium dodecyl sulfate (SDS) and 1 mm garnet rocks, followed by SDS cleanup with 1/3 volume 5 M potassium acetate, and then SPRI beads. The library was prepared using the TruSeq Nano kit (Illumina, San Diego, CA, USA), with DNA shearing performed with a S2 focused ultrasonicator (Covaris, Woburn, MA, USA) as suggested in the manufacturer’s protocol. Rather than Illumina adapters, we used custom adapters described in ref. [24 (link)]. The sample was sequenced on one lane of a HiSeq 2000 instrument (Illumina), using a 100 bp single-end kit.
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