I am trying to find a good way to isolate exosomes other than ultracentrifugation. Has anyone tried the Qiagen kit? Are there good methods that others would suggest?
A kit from SBI works well (see link below).
Link: https://www.systembio.com/microrna-research/exoquick-exosomes/overview
the Total exosome isolation reagents (5 for misc sample types) work really well
Both these suggestions- exoquick and total exosome isolation reagent- are PEG based, non-specific precipitation methods, and will isolate non-exosome contaminates as well
has anyone used the end products of exoEasy Maxi Kit (76064) for analyzing the RNA? Is it a good kit for such purposes?
I have used RNA extracted with exoeasy maxi and midi kit for NGS and QPCR.
It works, but we are not sure what types of EVs are captured, we can see EVs in the flowthrough. Not sure if it is that there were to many to bind or if other type of optimization is required. I have also extracted RNA from EVs prepared with exospin columns and they looked very homogeneous, I also used them for RNA extraction (using miElute RNA column from Qiagen) and NGS. RNA yield from gradient centrifugation is low so it works better if you have a good amount of starting material. EVs appear slighltly different as I saw no detectable rRNA 18 or 28s with ExoEasy, but there is some with gradient centrifugation and exospin. So take your pick. But best is to set up a method for your application keeping in mind that there are many different EVs and if cleanness and purity are an issue gradient centrifugation is still what will get you cleanest EVs, but you loose a lot.
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