RNA isolation / purification Yeast - Neurospora crassa

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 3 matching solutions for this experiment

Protocol tips
RNA was extracted and purified with the RNAprep Pure Plant Kit (Tiangen Biotech). Poly(A)+ mRNA was then isolated with the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) as described by the manufacturer.
TRIzol Reagent

Thermo Fisher Scientific

Protocol tips
For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.
Downstream tips
RNA was purified using a Qiagen RNeasy kit, and treated with Ambion Turbo DNase
Protocol tips
Both enzymatic and mechanical lysis can be used for yeast. Enzymatic lysis doesn't require additional lab equipment, but takes longer than mechanical disruption.
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