When I extract total RNA from serum specimens by Trizole or Roch kit the 260/280 OD are above 2(2.7) so the quality are not good for NGS. What is the best method to achieve the acceptable quality for NGS?
A great paper that compares 6 or 7 commercially available kits, many of which I've tested, is Burgos, K., Javaherian, A., Bomprezzie, R. et al. Identification of extracellular miRNA in human cerebrospinal fluid by next-generation sequencing. RNA. 2013 May;19(5):712-22. doi: 10.1261/rna.036863.112. Epub 2013 Mar 22.
The paper has nice data concerning each kits performance. I think you'll find it useful even though it is with CSF. Good luck.
Hi,
we have very good experience with Exiqon's products, as they are specialised in miRNA quantification from biofluids. Please take care, that not only the miRNA isolation is of importance but also the choice of library prep for NGS. NEB is good, Illumina is most common in application but I can not recommand that, because validation of quantitative differences by qPCR is often not possible.
There is another good publication you should look at (see attachement)
Dear Julian & Jeffrey
Thanks for your valuable answers. I will try more based on your anwers.
Best wishes
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