I have been isolating RNA from CD14 monocytes purified from PBMCs but the yield has been low. I've read that DNase treatment can result in lower RNA yields, but I want to ensure that these results can be published in the near future also.
Depending on the application of extracted RNA, DNase treatment could be essential. Certain RNA application are sensitive to very small amounts of gDNA contamination for example RT-qPCR. Quiagen RNeasy mini kits do not ask for DNase treatment since the silica-gel–membrane, spin-column technology efficiently removes the majority of the DNA without DNase treatment. On the other hand Aurum total RNA mini kit from Biorad supplies the lyophilized DNase to be used for gDNA removal from RNA and subsequent RT-qPCR.
Thanks for your answer Nupar - I'm using the RNA for qPCR, so perhaps a DNase treatment should be performed to ensure total removal.
In case of RT-PCR, DNase treatment is necessary, because it would eliminate gDNA contamination.
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