Many Thank´s! I already knew about this kit. But till now, I didn´t know if it really works.

oh really ?

I've heard that the columns of miRNeasy kit let a substantial quantity of miRNAs flow through...

What other kits did you try ?

regards.

Personally, I use the TRIZOL LS (for Liquid Samples) or mirVANA kit.

But still my best secreted miRNA isolations were performed with a exosome purification step prior to miRNA isolation itself.

How is that exosome purification?

It will depend on what you are working but if I am correct, in the litterature there are several population of secreted miRNAs. contained in exosomes (50-200 nm vesicles), microvesicles (>500nm vesicles) and free miRNAs. If you isolate the particles containing the miRNAs you will discriminate your miRNAs populations and improve the isolation. Hope this helps.

Hi Paula

Hope not to be too late... I am using the protocol described by Exiqon (see the link for a PDF file). These people from Exiqon are commercializing now a new kit for the extraction of miRNAs and other RNAs from biological fluids. I never used, but the feedback that I got from some customers is EXCELENT... Let me know if I can help.

Good luck.

Paco

http://www.exiqon.com/ls/Documents/Scientific/microRNA-serum-plasma-guidelines.pdf

You both really helped. Thank you!

Mirvana kit is good

I've always used miRVANA kit. Works pretty good and the advantage is that you can measure mRNAs in the very same RNA. Plus, miRVANA PARIS allows you also to extract protein from the same cells, so you can have miRNA, mRNA and protein from the same cells.

Martin, how do you extract proteins with miRvana kit? it's not mentioned in the protocol

Http://tools.invitrogen.com/content/sfs/manuals/cms_056986.pdf

Hi Assaf, please, see the link.

Hi Martin, I wasn't familiar with this kit, I work with mirVana miRNA isolation kit which doesn't have the option for protein extraction. Thanks.

Hi, I would just like to mention that several other products are available for both generic enrichment of exosomes as well as a bead based isolation of exosomes by targeting exosome surface markers such as CD63. Such bead bound exosomes can be used e.g. for downstream flow analysis, WB and TEM

http://www.invitrogen.com/search/global/searchAction.action?query=exosome&resultPage=1&resultsPerPage=15

kind regards

ketil