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.

Start discussion

No discussions found

Share your thoughts or question with experts in your field

Start a discussion

Found 3 matching solutions for this experiment

E.Z.N.A.® Fungal RNA Kit

Omega Bio Tek

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

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)

Manufacturer protocol Publication protocol 1 Relevant paper

RNeasy Plant Mini Kit

Qiagen

Upstream tips	Protocol tips	Downstream 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.

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.

Manufacturer protocol Publication protocol 1 Relevant paper

TRIzol Reagent

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Manufacturer protocol Publication protocol 1 Relevant paper

Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!

Fill out your contact details and receive price quotes in your Inbox