Welcome to ExosomesTalk, a new initiative for scientists in the exciting field of exosome research.

Join the conversations by responding to questions posted by your fellow scientists, and post your own questions.

ExosomesTalk is curated by scientists at Life Technologies. , with all content focused on your research needs. Our goal is to help you achieve faster breakthroughs by building products and services tailored to your needs.

Are there anyone isolate exosome RNA by TriZol reagent?

I cannot see the PCR product and RNA pellets after i collected serum-free media (only DMEM) 48 h. What should i do and any other method for that.
asked Jan 7, 2015 in ExosomesTalk by BiggieTanapong

2 Answers

Yes, I have done this.  Here is what I did:

  • re-suspended the exosome pellet in 250 ul PBS, transferred to a 1.5 ml microtube, added 750 ul Trizol LS.  It's important to use Trizol LS, which is specially formulated for liquid samples.  
  • Performed the RNA isolation according to the manufacturer's instructions with one important modification.  After the phase separation, when you have transferred the clear upper aqueous phase to a new tube, add 5 ul GlycoBlue (available from Ambion). It contains glycogen, which can help precipitate smaller amounts of RNA. This amount of glycogen (7.5 ug) doesn't inhibit cDNA synthesis or PCR.  Also it contains a blue dye that makes it easier to see the pellet.  

This increased my yields x10.  Good luck!

answered Jan 24, 2015 by KristinL

Another tip:

In the same RNA precipitation phase cited by KristinL I place the aqueous phase with Isopropanol overnight at 4°C. I saw that can icrease critically the RNA yelds.



Exosome Isolation for Proteomic Analyses and RNA Profiling, Douglas D. Taylor, Wolfgang Zacharias, and Cicek Gercel-Taylor; Methods in Molecular Biology, vol. 728, DOI 10.1007/978-1-61779-068-3_15, © Springer Science+Business Media, LLC 2011

answered Mar 9, 2015 by MBonetti
edited Mar 9, 2015 by MBonetti
thank you for u comment