Hello. Are you talking about universal primer for mIRNA analysis with real time PCR?

Just trying to identify universal primer for various pathogenic microbes detection, based on conserved motiffs

What you are telling please send me so i further read it and may be apply .

Thanks for your interest

Well i think you are on the right track. Just download the dna sequences corresponding to the conserved motifs. Primer should be 22-25 residues long. The 3 residues from the 3'end of the primers should be conserved in all the sequences. If variability exists in other sites, introduce a degeneracy at that site but not more than 2 degeneracies per primer (4 at one site or 2 at 2 sites).

Also since you want it for detection ideally keep the pcr product size between 300-500 bp. I am sure there are better ideas, but the above methods worked for me

Hi. I am sorry but I have no experience with this kind of analysis. I thought you wanted to design universal primers for miRNAs.

regards

Antonio

Thanks everyone for giving your valuable answers if any one worked on it kindly provide me your papers so i can go through it, either wet lab or in dry lab.