Hey guys,
Is it necessary to use the small RNA enrichment protocol to do miRNA study in this mirVana miRNA isolation kit? Cuz i found that using the total RNA extraction to perform real-time PCR quantifying miRNAs is totally fine. I know this kit receiving lots of good feedbacks and i ordered one. I tried using the small RNA enrichment protocol and the product concentration is pretty low, like 78ng/ul. And i added 100ul 95 degree pre-heat water to elute the miRNA off the column, so does anyone have any experience about this kit?
Dear Mr Dong, good afternoon.
I'm using this kit to extract miRNAs from kidney tissues, and it is pretty easy to do it.
For quantify miRNAs you don't need enrich the sample for small RNA, only extract the total RNAs is sufficient to quantify miRNA by qPCR.
As the amount of miRNA per microliter of sample, I usually use 20ng/ul, sometimes 10ng/ul and it is sufficient for me.
One thing that can help you, make a serial dilution using 3 or more different concentrations of a pool of samples in triplicate and compare the results to see what is the better concentration for your case.
I hope it help you.
Best regard
Pedro
i used this kit to isolate microRNA from cultured cells and i got very less concentration after elution. Qiagen kit is best for isolating microRNAs from my experiance
I used this kit and got pretty good product concentration from 200-1000ng/ul for total RNA extraction.
Does anyone know the chemical composition of elution solution?
Hi,
I am trying to isolate total RNA from mouse brain. I get around 140-180 ng/ul.
How to increase the yield? How to perform tissue disruption? I currently use mortar and pestle, semi automated; put the frozen tissue directly into lysis buffer.
Maybe if you lyophilize the sample you can increase the concentration Diji Kuriakose .
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