RNA isolation / purification Yeast - Candida albicans

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
For Candida albicans, a more specific protocol might be required (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"
Upstream tips
Samples can be flash frozen (-80C) before use, this will likely cause lower RNA yield.
Downstream tips
The isolated RNA was treated with RNAse-free DNAse I.

When checking the yield by means of a spectrophotometer, use the supplied TE buffer to zero the machine
Upstream tips
DNase treatment and RNase inhibitor are included in this kit
Downstream tips
RNA was treated with DNase (Epicentre) to remove contaminating genomic DNA.
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