Hi all!
I was wondering if anyone has experience or information about storing whole blood samples in RLT buffer for future RNA extraction. In my case, I will need to process/store a lot of blood pellets after plasma collection at the same time, and isolate PBLs of this amount of samples will be complicated.
I know that this RLT buffer is a lysis buffer itself, but I am a little bit concerned about the quality of the RNA for performing RNAseq.
Thanks a lot for your time and suggestions!
JC.
Hi Juan Camilo Sánchez-Arcila ,
I have never done it with Qiagens RLT buffer, but you can lyss cells in TriZol and store frozen until use with high quality yield. Since I have never done it with RLT, I would try contacting qiagen technical support to see what they say. Otherwise try and experiment. Process fresh and process a sample that was frozen and run a denaturing gel to see if there is any degradation.
Hello,
I haven't tried storing samples for RNAseq in RLT buffer. However, I have done RNAseq for de novo transcriptome assembly from samples stored in RNAlater (Thermo Fisher). I endorse the quality of the RNA stored on this solution, it is compatible with TRIzol as well as with Qiagen or other kit extraction and the bottle is not too expensive. Besides, after processing you can even transport the samples at room temperature. I would definitely recommend RNAlater for that.
Hope this helps.
Annia
Hi, this article can help
https://pdfs.semanticscholar.org/4950/69834049f556a065577869972b439ce76598.pdf
If you can collect two tubes one for plasma and one only for RNA extraction for whole blood, maybe you can use these tubes
https://www.qiagen.com/ca/products/diagnostics-and-clinical-research/clinical-sample-processing/paxgene-blood-rna-tube-ivd/#orderinginformation
Hi Annia, were your samples whole blood!? And thanks Shawn Krueger , Annia Alba and Valeria De Arcangelis for your comments :)
Hi Juan Camilo,
Concerning your more recent question, no, my samples were not whole blood, it was other animal tissue. It says in the guidelines that can be used with whole blood but I have no particular experience with RNAlater and whole blood.
After a very quick search, I found a similar discussion in research gate that might be helpful (see the link: https://www.researchgate.net/post/Does_anyone_use_RNAlater_for_storing_blood_samples). In any case you can always run some tests with the different extraction methods by analyzying the qualitity of the extracted RNA on a Bioanalizer.
Good luck!
Annia
Hi Juan Camilo Sánchez-Arcila :D
Check number 3 on the RNA later FAQs:
https://www.thermofisher.com/se/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rnalater-faqs.html
Un abrazo :D
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