RNA isolation / purification Yeast - Schizophyllum commune

Generally it has been difficult to isolate high-quality RNA from yeast because of problems disrupting the cells. Use of enzymes to disrupt cell wall can alter gene expression profiles. Therefore, physical disruption can result in high quality RNA for all downstream processing. Use of DNAse and proteinase K will remove traces of DNA contamination and proteins respectively.

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Found 2 matching solutions for this experiment

RNeasy Plant Mini Kit

Qiagen

Upstream tips	Protocol tips	Downstream tips
	RNA of seven days old, solid cultures of the strains 12–43, E6 and ∆dhc2 was isolated using RNeasy Plant Mini Kit performing an additional DNAse digestion with RNase-Free DNase Set (both Qiagen, Hilden,Germany). RNA-sequencing was performed by LGC Genomics, Berlin (Germany) with mRNA-based cDNA-libraries constructed from sequencing adaptors ligated to cDNA fragments

Protocol tips
RNA of seven days old, solid cultures of the strains 12–43, E6 and ∆dhc2 was isolated using RNeasy Plant Mini Kit performing an additional DNAse digestion with RNase-Free DNase Set (both Qiagen, Hilden,Germany). RNA-sequencing was performed by LGC Genomics, Berlin (Germany) with mRNA-based cDNA-libraries constructed from sequencing adaptors ligated to cDNA fragments

Manufacturer protocol Publication protocol 1 Relevant paper

TRIzol Reagent

Thermo Fisher Scientific

NucleoSpin® RNA

Macherey Nagel

Upstream tips	Protocol tips	Downstream tips
	Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Protocol tips
Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Manufacturer protocol Publication protocol 1 Relevant paper

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