2 replies to this topic

[epibet]

Hi guys!
I have isolated RNA from very precious frozen samples by using Trizol, but i have DNA contamination which i found out by using no RT. Is there any DNase treatment protocol i can use to eliminate the DNA from my RNA samples with minimum RNA loss. Thanx.

[Curtis]

Certainly, there are plenty of protocols on Protocol-online.org but it depends on the company protocol as well. I mean the commercial DNase from different companies might have different protocols. My DNase is from Fermentas and it came with a stop solution to stop the reaction after a certain time.

But I don't understand how you figured out you have DNA in your sample. no RT? what is that?

When extracting RNA from Eukaryotic sample you would normally end up with DNA contamination.

[epibio]

I believe epibet is referring to using the RNA as template for RT-PCR. When you do a control without reverse transcriptase (no RT) and your PCR still gives you positive results, it would indicate DNA contamination.