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mirVana™ qRT-PCR miRNA Detection versus miScript SYBR Green PCR kit - advantages and disadvantages (Nov/09/2008 )

I’m just starting to work with miRNA and would be very grateful if you could give me some help. I’m looking for a method for detecting mirRNA by PCR and I have read about: mirVana™ qRT-PCR miRNA Detection Kit Primer Sets and miScript SYBR Green PCR kit from QIAGEN. I have read some comments about the first one, “mirVana”, in this forum and the majority of people recommend the use of Taqman instead of primers. Moreover, I don’t like the fact of doing 2 separate RT reactions for detecting my miRNA and a housekeeping gene. However, I haven’t found any comment about miScript SYBR Green PCR kit from QIAGEN. I would like to know if someone has used it and if it works correctly. Any advice on which one to use ????? huh.gif

Thanks in advance !! rolleyes.gif
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-pilar-

You should think about normal isolation of total RNA by Trizol as many people claim it is easier and better than using a kit. I used miscript, isolation and RT worked fine, i just have got efficiency problems in the pcr, but that may be because of other problems. I think the advantage of miscript is just that during RT there will be added an anchor for a universal primer on all miRNAs in the sample, so subsequently you can analyse different miRNAs out of one sample.
What do others say?

-mirno1-