I have prepared the RNA ladder twice from two different stocks. It is prepared in eppendorph tubes from the same box as I prepare my RNA samples in, and the RNA samples look normal and fine on the Bioanalyzer. I think it must be something with the ladder. Do you have any suggestions to what is wrong and how I can fix it?
My ladder gave me the same picture. I have no idea what's wrong. First I thaught the ladder was diluted with too much water but it turned out that everything was done correctly. Now I'm clueless.
Did you store the ladder on dry ice during transport and store it at -80 after use? It might be degraded.
I keep the diluted and aliquoted ladder tubes in the -80 freezer. It is a relatively newly bought ladder and I received two stocks. First I prepared one of them while keeping the other in the -20 freezer. When I realized that it looked strange on the Bioanalyzer, I prepared the other stock and the result is exactly the same on the Bioanalyzer. I highly doubt that it has been degraded both times, that I have prepared the diluted ladders as I did it two different places and used RNase-free water from two different flasks. It is so strange and annoying!
Make sure you aliquot the ladder into the provided low-binding 0.5 mL tubes. I reran with diluted ladder taken straight out from the original ladder tube and all the peaks were resolved except for one.
Hi all,
We had exactly the same problem. The ladder was not degradated. In fact, it seems that the problem was just the syringe clip which was set at the lowest position (as for High sensitivity chip)...
I haven't been able to resolve the problem but I also did not try the last 6 months. Thanks to Veronique for that suggestion, I don't recall which position I had the syringe clip in, but I will look into that next time! Glad you solved the problem in your lab :-)
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA