Am used blibirt kit for isolation circulating micro RNA that included three steps

@Mutta what's the yield of MiRNA? And what types of kit you used for cDNA?!

Hi there

I have experience with miRNeasy Serum/Plasma Kit, which is a modification of the miRNeasy kit devoted for biofluids.

For qPCR, I have experience with the Exiqon system with LNA primers. For this protocol you can use any other system, but be sure to employ the proper cDNA synthesis kit, since classical protocols for cDNA synthesis will not work with miRNAs because of their size.

Best of luck

@Francisco

Thank you for your reply

hi,

We have not used the particular Qiagen mirneasy kits. We are doing a miRNA study in cancer cells and mesenchymal stem cells. There we isolated total RNA from cells using the usual TRIzol method.

Then as Francisco J. Enguita mentioned about critical step is of cDNA synthesis. Here, we used the Qiagen miScript II RT Kit which converts all miRNAs in total RNA into miRNA-specific cDNA. That cDNA can be used directly for qPCR using SYBr green. Also, Qiagen pre-synthesized LNA primers specific to miRNA are being used in qPCR. I am just sharing an approach, which may be applied to your study also.

Regards

Saurabh

@Saurabh Mendal. thanks, OK I'll try minScript kit for cDNA.