title: GSM4065471: ∆lsd1 N5555 RNA-seq rep1; Neurospora crassa; RNA-Seq
library_construction_protocol: Cultures were grown in Vogels medium and 1.5% sucrose (standard minimal medium), shaking for 16 hours at 32˚C. Mycelia were harvested and added to a tube containing ~350µL glass beads (260-300 micrometer, acid washed [Sigma Aldrich] and resuspended in sdH2O), 350µL NETS buffer (10mM Tris pH 7.5, 300mM NaCl, 1mM EDTA, 0.2% SDS) and 350µL phenol:chloroform:isoamyl alcohol (25:24:1) and lysed by a bead beater for 3 minutes at room temperature. Samples were cleared by centrifugation, and the aqueous (top) phase was mixed with 350µL of chloroform. After centrifugation, the aqueous phase was divided into two tubes containing 650µL cold 100% EtOH.  Samples were precipitated at -20˚C, pelleted by centrifugation, washed twice with 1mL 70% EtOH, and resuspended in DEPC-treated dH2O. RNA was treated with DNAseI, amplification grade (Thermo Fisher Scientific), cleaned (Agencourt® RNAclean XP® beads, Beckman Coulter), and RNA-seq libraries were prepared (KAPA Stranded mRNA-seq kit, KAPA Biosystems).
attributes.tag: GEO Accession
attributes.value: GSM4065471