RNA isolation / purification Yeast - Aspergillus nidulans

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

Upstream tips
Cell disruption was performed in a cell homogenizer (Retsch MM200) with glass beads (Qiagen) at 30 hits s−1 for 3 min.
Protocol tips
If RNA remains on the column, heat DEPC water to 65 °C prior to elution
Downstream tips
The isolated RNA was quantified and treated with TURBO DNA‐free kit (Ambion)
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.
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.
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