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[Neanderthal]

Hello everyone,


This is already posted on miRNA discussion group. But I am resposting it because of its relevance. Please bear with me.

I am interested in doing qRT-PCR to validate my miRNA microarray analysis. In order to do miRNA qRT-PCR,

1, How do I design primers for my selected miRNAs?

2. Should I take mature or stem loop structure?

3. Is there any software avalibale for designing miRNA qRT-PCR primers?

4. Do we need any special chemicals/reagents for miRNA qRT-PCR other than the standared reagents used with SYBR green chemistry for normal qRT-PCR (by normal I meant qRT-PCR for genes like p53, GAPDH etc)

5. What should I use as my housekeeping gene in case of miRNA-qRT PCR?

If someone can provide a guidline for desiging miRNA-qRT PCR primers, it will be of immense help.

Thanks a lot in advance.

K.