Hello everyone, I´m using the Agilent's Small RNA chip for detection of microRNA, but I´m not sure of the indicated concentration by the bioanalyzer (18% of miRNA), because I didn't observe the "hill like" pattern showed on the manuals (fig 1), instead I observe "peaks" (fig 2) on the miRNA region (from 15 to 40 nt). Is that enough evidence of the presence of miRNA in the sample?
Hi,
your run looks little weird indeed. I never had dedicated peak for miRNAs but a shoulder. It looks like you have some issues with the chip because it looks so staggered and not consistent. What samples is it?
I'm not sure how pure your sample and what kind of method are you using for RNA purification. I would make sure your RNA is pure and dissolved in ultra pure water .
Looks like a bad run to me. The marker (at 4nt) is barely detected. How did the ladder & other wells look? If there's any surfactant in one of the samples, that can ruin runs for the entire chip.
Thanks everyone a lot for your answer and invaluable information; this results are part of a method standarization, so it still needs some fitting but i want to be sure that at least I was isolating miRNA´s. In fact the obtained ratio 260/230 was lower than recomended and that could cause the baseline going up so now I know that I have to be more carefully with this parameter
- Badges
- Science topic
- Similar topics
- Chemistry
- Biochemistry
- Biomolecules
- Nucleic Acids
- RNA
- Small RNA