I extracted RNA from liver tissue from fish using the trizol protocol.
When running a gel, I get what looks like a high molecular band at the top of the gel, within the well, that doesn't move into the gel.
I used post-gel staining with Gel Green.
Does anyone have an idea what it could be? I was thinking it is either gDNA or protein. But I wouldn't have thought that Gel Green binds to protein.
I treated the RNA with DNase and ran another gel, but the band was still there.
Hi, maybe this will help you: https://www.researchgate.net/post/What_could_cause_RNA_bands_to_not_migrate_on_a_denaturing_agarose_gel
Cheers, Nadine
Enjoy diving :-)
Thanks, the only answer that could fit is the suggestion of protein/RNA complexes that don't migrate. Although the nanodrop values weren't that bad to suggest this. It's certainly not a dye artefact since I post-stain my gels.
Hi Roger,
I think your “high” molecular weight bands could have different reasons that need different strategies to solve them:
If the bands are pure aggregates try to dissolve your RNA in RNase free buffer (10-20 mM, pH 7-8) and may heat the samples to 65°C if necessary.
If a protein contamination causes aggregation try to include a proteinase K treatment in your preparation.
Sometimes loading loading buffers containing urea or formaldehyde or formamide got too old and cause modification of RNA as well as electrophoreses artifacts. Prepare a new loading buffer with fresh de-ionised reagents.
Best,
Bodo
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Proteins