I encounter the following problem: I want to send viral RNA for further analysis. As the receiver requires, I must process my samples according to a regular RNA extraction protocol and send the filtered spin columns from the RNA extraction kit, without performing the final elution step. But I don't have the original sample preserved, only the RNA already extracted is stored. Can I do a re-extraction using the stored eluate as the starting material?
Hi there,
Sure you can provided you apply the conditions required for the efficient binding of RNA to the matrix.
Yes you can! Like Dominique said, make sure to mix your already eluted RNA in the binding solution (eg. Trizol and Ethanol mix) and then load the column for successful binding.
Thank you all. My great concern is that I use an extraction protocol, according to which: first, I have to bring the original sample to a volume of 200 microliters (I am afraid that I will dilute the sample too much), and second, I have to mix the sample with a lysis buffer, containing carrier RNA and then to incubate the mixture for 15 minutes at 56 C. As far as my starting material is pure RNA, I am concerned that this step could damage the RNA.
Hi Irina Georgieva
I believe that, even if your RNA sample is pure, making up its volume to 200 µL will be OK because most columns have high affinity for RNA. No doubt, you will lose some RNA, but I hope it will be enough for you analysis.
Secondly, don’t worry about RNA degradation at 56°C for 15 minutes because we routinely perform proteinase K treatment at 55°C overnight to separate RNA from proteins. Secondly RNA is exposed to 70°C for 10 minutes for melting before cDNA synthesis. RNA is stable despite exposing to these temperatures.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA