title: SPRs-PN1-2-Rep3
organism: Solanum tuberosum
description: Protocols: 3 and 8 days after innoculation samples were taken from the sprouts. Samples were directly froozen in liquid nitrogen. Sprouts of ten plants per replicate were sampled and pooled. Samples are denominated as following: SPK-PN1-x-Repx and SPK-PN2-x-Repx stands for sprouts without pathogen inoculation sampled at 3 and 8 dpi and SPRs-PN1-x-Repx and SPRs-PN2-x-Repx indicate sprout samples with Rhizoctonia inoculation at 3 and 8 dpi. For the R. solani kontrol 3 samples were extrated from plates grown on PDA agarose. Potato cultivar Arkula tubers (Norika GmbH, Sanitz, Germany) were grown on a cycle of (1 week 8 degrees Celcius, 3 weeks 4 degrees and 3 days 20-22 degrees), until sprouting. 10 mg/L gibberellic acid was applied for 20 minutes. The tubers were grown in 12x12x10 cm pots with quartz sand and grit mixture in the dark at 18 degrees and 80% relative humidity. R. solanig AG3 isolate Ben3 was cultivated on PDA agar plates (PDA, Merck, Germany) at 20-22 degrees Celcius for 5 days. 4 Weeks after transplantation into pots the tubers were innoculated with a 1cm agar plug from the R. solani plates and the tubers were covered with quartz sand. 3 and 8 days after innoculation samples were taken from the sprouts. Each sample was ground using a mixer mill (2 min, 30/s; Retsch MM400, Haan, Germany) with two grinding balls (7 mm, 3 mm; Askubal, Korntal-Münchingen, Germany) under con-stant cooling in liquid nitrogen. Total RNA was extracted from 70-90 mg of ground sprout material (without lesions) using the RNeasy Plant Mini Kit (QIAGEN, Hilden, Germany) including DNase treatment (QI-AGEN). From each biological replicate 3 technical replicates were generated resulting in a total of 36 libraries with the TruSeq® mRNA Sample Preparation Kit (stranded) (Illumina Inc., San Diego, U.S.A.).