title: An improved and reproducible protocol for the extraction of high quality fungal RNA from plant biomass substrates.
abstract: Isolation of high quantity and quality RNA is a crucial step in the detection of meaningful gene expression data. Obtaining intact fungal RNA from complex lignocellulosic substrates is often difficult, producing low integrity RNA which perform poorly in downstream applications. In this study we developed an RNA extraction method using CsCl centrifugation procedure, modified from previous reports and adapted for isolation of RNA from plant biomass. This method provided high level of integrity and good quantity of RNA which were suitable for reliable analyses of gene expression and produced consistent and reproducible results.
authors.affiliation: Fungal Physiology, CBS-KNAW Fungal Biodiversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands; Fungal Molecular Physiology, Utrecht University, Utrecht, The Netherlands.
authors.forename: Aleksandrina
authors.initials: A
authors.lastname: Patyshakuliyeva
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki, P.O. Box 56, 00014 Helsinki, Finland.
authors.forename: Miia R
authors.initials: MR
authors.lastname: Mäkelä
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki, P.O. Box 56, 00014 Helsinki, Finland.
authors.forename: Outi-Maaria
authors.initials: OM
authors.lastname: Sietiö
authors.affiliation: Fungal Physiology, CBS-KNAW Fungal Biodiversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands; Fungal Molecular Physiology, Utrecht University, Utrecht, The Netherlands. Electronic address: r.devries@cbs.knaw.nl.
authors.forename: Ronald P
authors.initials: RP
authors.lastname: de Vries
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki, P.O. Box 56, 00014 Helsinki, Finland.
authors.forename: Kristiina S
authors.initials: KS
authors.lastname: Hildén
mesh_terms: D018533:Biomass; D002499:Centrifugation, Density Gradient / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods; D008967:Molecular Biology / Q000379:methods*; D010944:Plants / Q000382:microbiology*; D012331:RNA, Fungal / Q000302:isolation & purification*
chemical_list: D012331:RNA, Fungal
keywords: Agaricus bisporus; CsCl gradient ultracentrifugation; Dichomitus squalens; Gene expression; Lignocellulose (plant biomass); RNA extraction