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Isolation of Total RNA From Neurospora Mycelium

Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 362)

Abstract

In filamentous fungi, including the model organism Neurospora crassa, plentiful biological tissue from which RNA can be extracted may be obtained by allowing fungal spores to germinate and form a mycelium in liquid culture. The mycelium constitutes a mosaic of multinuclear, tubular filaments known as hyphae or mycelia. In general, when exposed to air, fungal hyphae quickly start to develop spores, which are often colorful. However, when submerged in liquid under rapid agitation large amounts of vegetatively growing mycelium can be obtained, which can be easily harvested by means of filtration. To preserve the physiological state of the culture, the mycelium is snap-frozen, and then to free its contents, the mycelium is ground under liquid nitrogen to break all hyphal structures. Here a method to extract high-quality total RNA from Neurospora mycelium using TRIzol® reagent is described.

Key Words

Circadian filamentous fungus bread mold hyphae mycelium mycelial disk liquid nitrogen RNA TRIzol 
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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  1. 1.Department of GeneticsUniversity of LeicesterLeicesterUK

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