I am using the Thermo Scientific DNase I (https://www.thermofisher.com/order/catalog/product/EN0521) to remove genomic DNA from RNA extracts eluted in nuclease free water.

The protocol calls for 1 u of DNase enzyme for 1µg of RNA, so a typical reaction would look like 8µL of sample, 1µL of 10x buffer, and 1µL of 1u of enzyme for a total rxn volume of 10µL. This protocol is typical for other DNase I treatment kits that I have come across.

Problem is, I have a handful of samples that are low concentration (i.e. less than 1µg of RNA in the suggested 8µL of sample). If i was using another kit, it seems like it is okay for DNase I enzyme to be in excess of the nucleic acids. However the brochure for this product has the line "Do not use more than 1 u of DNase I, RNase-free per 1 µg of RNA."

How should i proceed?

Should I just use 8µL of sample regardless?

Should I be tailoring the amount of enzyme i use to each reaction so the ratio of u enzyme to µg of sample is always less than 1? (if I do the second option do I need to adjust the amount of Buffer and EDTA as well?

Im inclined to go with option based on other results on the internet

More Isaac Vingan's questions See All
Similar questions and discussions