RNA isolation / purification Yeast - Cryptococcus neoformans

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

RNeasy Mini Kit

Qiagen

Upstream tips	Protocol tips	Downstream tips
	C. neoformans might require extra effort for lysis (bead beating, see reference) To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	"- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C"

Protocol tips
C. neoformans might require extra effort for lysis (bead beating, see reference) To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
Downstream tips
"- Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C"

Manufacturer protocol Publication protocol 1 Relevant paper

PureLink™ RNA Mini Kit

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
For lysis of yeast, extra enzymatic or mechanical disruption is recommended.	Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required	DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit

Upstream tips
For lysis of yeast, extra enzymatic or mechanical disruption is recommended.
Protocol tips
Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required
Downstream tips
DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit

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

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