Hi
I have a question about protocol of isolation RNA by Mirvana microRNA kit. I want to detect expression mirna in rats tissue of mammary gland tumor using of Taqman mirna. Is it correct use total RNA isolated from tissue or I must use protocol for small RNA enrichment. I found in articles that using of total RNA is sufficient
Thanks
Dear Peter Observnik,
I used this same kit to do my research during my doctorate when I began I tried to do the extraction of miRNA by the protocol doing the enrichment step, and I could not see amplification. So, I asked about this issue to the scientific advice from Life technologies, who sold this kit for us, and they told me that I must do the extraction only until the total RNA without small RNA enrichment. So if you do in this way you will get the expression.
I hope that it helps you.
Pedro
Total RNA purification protocol works fine; you can optionally enrich for small RNA. qRT-PCR with miRNA TaqMan work great on RNA purified with mirVana kit (there are spin column based options and magnetic bead based options for high throughput)
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