Citation:
Goncalo M. Leite, Naresh Magan, Angel Medina. Comparison of different bead-beating RNA extraction strategies: An optimized
method for filamentous fungi. Journal of Microbiological Methods, 2012, Volume 88, Issue 3, pp413-416.
Abstract:
Molecular studies, especially in relation to the activity of secondary
metabolite gene clusters, require the ability to extract good quality RNA from
fungal biomass. This is often hindered by the cell wall structure and endogenous
RNase activity in filamentous fungi. There is thus a need for rapid methods for
the extraction of good quality RNA for use in microarrays and for quantitative
PCR assays. The objective of this study was to examine the use of different
systems for the high throughput method to extract intact RNA from filamentous
fungi. Two bead beating systems with different motion patterns and speed
capacities were tested in the development of the extraction protocol. They were
evaluated based on the total RNA yield and overall RNA quality. The high speed
bead beating with glass beads associated with an automated purification method
gave more than three times higher total RNA yields with less than a quarter of
the amount of mycelium required. Furthermore the integrity and overall quality
was conserved, with RNA Quality Indicator (RQI) numbers consistently >7.5. This
method also reduced cross contamination risks and kept RNA handling to a minimum
while still being capable of multiple sample processing, reducing the time
required to obtain RNA from filamentous fungi.