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 ?????
Thanks in advance !!
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?