title: Pycnoporus sanguineus transcriptome
design: Pycnoporus sanguineus strain 2126 (BAFC: Mycological Culture Collection of the Department of Biological Sciences, Faculty of Exact and Natural Sciences, University of Buenos Aires) (Polyporaceae, Aphyllophorales, Basidiomycetes) was cultured in GA medium  : glucose, 20 g; asparagine monohydrate, 3 g; MgSO4 ·7H2O, 0.5 g; KH2PO4, 0.5 g; K2HPO4, 0.6 g; MnCl2 ·4H2O, 0.09 mg; H3BO3, 0.07 mg; Na2MoO4 ·H2O, 0.02 mg; FeCl3, 1 mg; ZnCl2, 3.5 mg; thiamine hydrochloride, 0.1 mg, supplemented with 1 mM copper sulfate; distilled water up to 1 L. Initial pH of the medium was adjusted to 6.5 with NaOH 1 N. Erlenmeyer flasks (500 ml size) with 50 ml of medium were inoculated with four 25-mm2 surface agar plugs from a 7-day-old culture grown on malt agar (1.3% malt extract, 1% glucose, agar 2%). Incubation was carried out statically at 28 ±1°C. Cultures were harvested at day 21 for total RNA extraction.   Fungal mycelium was filtered and immediately ground into fine powder using liquid nitrogen. Total RNA was extracted using RNAzol RT reagent (Molecular Research Center Inc., Cincinnati, USA) according to the manufacturer´s instructions. The quantity of RNA was estimated by Nanodrop ND-1000 spectrophotometer (Nanodrop Technologies) and RNA quality was determined by formaldehyde RNA gel electrophoresis.   Poly (A) RNA was purified from total RNA using Dynabeads oligo (dT) magnetic beads (Invitrogen Life Technologies, Carlsbad, USA) and mRNA was broken into fragments of 50 to 2000 nucleotides by adding fragmentation buffer. Fragmented mRNA quality was assessed by Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). Short mRNA sequences were used for double strand cDNA synthesis using the cDNA Synthesis System Kit (Roche) and random primers, followed by purification by QIAQuick PCR Purification kit (Qiagen Inc., CA, USA). The final cDNA library was constructed using the GS FLX Titanium Rapid Library Preparation Kit (Roche).