Frozen mycelia powder was resuspended in a peqGOLD TriFast mix (PeqLab, catalog no. 30-2010), and RNA was extracted following the manufacturer’s instructions. RNA pellets were dissolved in an appropriate volume of RNase-free water [supplemented with RNase inhibitor (0.4 U/μl) (RiboLock, Thermo Scientific, catalog no. EO0384)]. RNA concentration was measured using NanoDrop (PeqLab).
Peqgold trifast mix
Manufactured by Avantor
PeqGOLD TriFast mix is a ready-to-use reagent for the isolation of high-quality RNA from a variety of biological samples. It is based on a mono-phasic solution of phenol and guanidinium isothiocyanate, which is commonly used for RNA extraction.
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2 protocols using peqgold trifast mix
1
RNA Extraction from Frozen Mycelia
2
Fungal DNA and RNA Extraction Protocol
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For isolating DNA, mycelium was scratched off a PDA plate with a spatula and DNA was extracted according to a quick extraction protocol [67 (link)]. Biomass samples for RNA extraction from liquid cultures were filtered with a Miracloth filter, shock frozen with liquid nitrogen and stored at − 80 °C. For the RNA isolation the peqGOLD TriFast mix (PeqLab) was used according to the protocol. The RevertAid H Minus First Strand cDNA Synthesis Kit (Fermentas) was used for cDNA synthesis. All qPCRs were performed with the Luna Universal qPCR Master Mix (NEB). Results were evaluated with REST 2007 [68 (link)] freeware by QIAGEN. All primers for qPCR are listed in Additional file 2 : Table S3.
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