title: GSM4304754: WT_NH4_rep3; Aspergillus nidulans; RNA-Seq
library_construction_protocol: Mycelia were strained and blotted dry before snap freezing in liquid nitrogen. Samples were stored at -80ºC for up to 12 months. Total RNA was isolated by grinding mycelia under liquid nitrogen and subsequent addition to RNA extraction buffer (7.0 M Urea, 100 mM Tris-HCl pH 8.0, 10 mM EDTA, 1.0% sodium dodecyl sulfate) followed by two phenol-chloroform-isoamylalcohol extractions and one chloroform extraction (Sambrook and Russell, 2001). RNA was precipitated in 3M ammonium acetate and 50% isopropanol, resuspended in DEPC-H2O, and re-precipitated overnight in 4M lithium chloride at -20ºC. RNA quality was determined by visualization after electrophoretic separation in a 1.2% agarose gel containing 1.1% formaldehyde run in 1x MOPS buffer (20 mM MOPS pH 7.0, 5 mM sodium acetate, 1 mM EDTA). RQ1 DNase (Promega) treatment of RNA followed the manufacturer's instructions. RNA was indexed with the TruSeq Stranded Total RNA Sample Prep Kit (Illumina) and fragmented to 180-bp. Libraries were sequenced using 50-bp single-end reads on the Illumina Hi-Seq 2500 platform
attributes.tag: GEO Accession
attributes.value: GSM4304754