Hey!
I am planning to extract RNA from my samples and subsequently remove any genomic DNA contamination using DNase I from Thermo Fisher (#EN0521 https://www.thermofisher.com/order/catalog/product/EN0521). I have attached a photo of the protocol I will follow for reference. After the DNase treatment, I will proceed with cDNA synthesis.
Considering that my samples may have varying RNA concentrations, can I prepare a single DNase I mix that I then distribute among the different samples? Should I base the volumes on the sample with the lowest concentration, or is there a more efficient approach? How should I do this?
Thank you for your insights!
Base the amount of DNase on the most concentrated sample to be sure that there is no DNA left after the tratment in any of your samples. DNase will not touch your RNA. One master mix (i.e. the same amount of DNase per sample, regardless of the concentration) should be enough - the amount of DNase should be saturating.
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