I have collected blood in Tempus™ Blood RNA Tube from Thermofisher to purify RNA and do whole RNA sequencing later on. After trying different kits and getting very low yield, I went for the "MagMAX™ for Stabilized Blood Tubes RNA Isolation Kit, compatible with Tempus™ Blood RNA Tubes" which is meant to work specifically with those tubes. I was surprised when I isolate some RNA getting the following: -0.1 ng/ul, 260/280=0.19, 260/230=0.00 1.4 ng/ul, 260/280=122.49, 260/230=0.00 1.9 ng/ul, 260/280=5.68, 260/230=0.01 0.3 ng/ul, 260/280=-1.78, 260/230=0.01 These are really weird results. The manual says you can isolate up to 6 to 25 μg of RNA from 3 mL blood, with 260/280ratio > 1.9. Did anyone worked in the past with the same combination of tubes/kit with similar troubles? Any suggestions on what could be the cause? Thanks in advance.
I haven't worked with these particular tubes but I've used PaxGene tubes before. We had to use a carrier to get our RNA, but we were looking for microRNA so that's a bit different.
I found a couple of articles that you've probably already seen:
Article Long-term storage of blood RNA collected in RNA stabilizing ...
Article Comparison of blood RNA isolation methods from samples stabi...
You should check your method against what these researchers did. How old was your kit? How long and how did you store the tubes before you extracted the RNA? All of these variables will ultimately give you lower yields. In terms of the quality it might be best to run the samples on a bioanalyzer because your ratios look all weird, however with a yield that low I'm not surprised. Running a bioanalyzer chip will give you a good indication on the quality of the RNA and is more precise than a nanodrop.
Thanks for your answer Sophie. I will definitively check those papers out. I will go through to process as probably there was something wrong in it.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA