A Tool Set for the Genome-Wide Analysis ofNeurospora crassaby RT-PCR
Year: 2015
Type: article
Abstract: Neurospora crassa is an important model organism for filamentous fungi as well as for circadian biology and photobiology. Although the community-accumulated tool set for the molecular analysis of Neurospora is extensive, two components are missing: (1) dependable reference genes whose level of expression are relatively constant across light/dark cycles and as a function of time of day and (2) a catalog of primers specifically designed for real-time PCR (RT-PCR). To address the first of these we have identified genes that are optimal for use as reference genes in RT-PCR across a wide range of expression levels; the mRNA/transcripts from these genes have potential for use as reference noncycling transcripts outside of Neurospora. In addition, we have generated a genome-wide set of RT-PCR primers, thereby streamlining the analysis of gene expression. In validation studies these primers successfully identified target mRNAs arising from 70% (34 of 49) of all tested genes and from all (28) of the moderately to highly expressed tested genes.
Source: G3 Genes Genomes Genetics
Authors: Jennifer M Hurley, Arko Dasgupta, Peter Andrews, Alexander M Crowell, Carol Ringelberg, Jennifer J Loros, Jay C Dunlap
Institutions: Dartmouth College, University of Pennsylvania
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,709
Awards: Functional Analysis and Systems Biology of Filamentous Fungi, Identification and Analysis of Circadian Clock-Controlled Genes, Genetic and Molecular Dissection of the Neurospora Clock, G4008756567, G6898742010, Genetic and Molecular Dissection of the Neurospora Clock, G8635067024, Genetic and Molecular Dissection of the Neurospora Clock
OpenAlex ID: W2140237914