Hi. In an attachment I am sending picture of a gel after my RNA isolation. Since I do not have any experience with RNA isolation I am somewhat confused in interpreting what could possibly happen to my samples. Basically my only question is – is my RNA degraded (before and after DNAse treatment, see the picture)? The concentration of my samples is approx. 200 ng/ul and I loaded 2 ul of the sample in each lane. What is especially puzzling me is when my samples are compared to the control (RNA isolated by my colleague from the same organism but after different treatment) the 5S band is much stronger in my samples. Could it be for example that the degradation products of my RNA are mixed with 5S RNA and therefore corresponding bands have stronger intensities? Because when I went through the pictures of degraded RNA available on the internet they do not look the same. Or could it be consequence of the treatment of my bacterial cells (higher Mn2+ concentration)?
Thank you guys for your answers in advance!
Yes, It looks degraded. Make sure you are using high quality DNase. I have had the same problem, my RNA bands disappeared after I gave the DNase treatment. I suspected my DNase to be contaminated with RNase. The gel does not look very good, try the Bleach gel (as referred by Maxim), it did work very good for me.
The gel picture attached herein was run on a bleach gel.
I hope it works for you.
Good luck
Jan, as far as I see you have lost your high molecular weight RNA - look at the paper attached for the ideology and a simple approach.
Article Bleach Gel: A Simple Agarose Gel for Analyzing RNA Quality
Yes, it looks like it is degraded. You should see two bright, large molecular weight bands from the rRNA in each of your wells, like you see in your far right well. RNA is fragile, but not super fragile. Make sure your reagents are fresh, clean your work space and store your samples quickly and properly.
Yes, It looks degraded. Make sure you are using high quality DNase. I have had the same problem, my RNA bands disappeared after I gave the DNase treatment. I suspected my DNase to be contaminated with RNase. The gel does not look very good, try the Bleach gel (as referred by Maxim), it did work very good for me.
The gel picture attached herein was run on a bleach gel.
I hope it works for you.
Good luck
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology