I am looking for a standard protocol of how to design a good primer for RT-PCR.
Could anyone recommend me the standard protocol of the design of (microRNA or gene) primers for RT-PCR as a reference?
Sokviseth Moeng
Varkonyi-Gasic and Hellens, (2011). This article gives you an idea to design primers for miRNA. I used the same strategy for the screening of virus as well as plant miRNAs. It worked.
Check this out
Varkonyi-Gasic, E., Wu, R., Wood, M., Walton, E. F., & Hellens, R. P. (2007). Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant methods, 3(1), 12.
Hope it helps.
If you are looking for miRNA primer, then prefer pre-made primer from company. miRNA primer have universal primer sequence, upon which company synthesize with little chemical modification to for stability and to enhance specificity.
Rest gene primer can be synthesized using primerblast..
Ramamoorthy Sankaranarayanan Thanks for your recommendation. Is it also good to read if we study animal microRNAs?
Agi Vivian Thanks for your recommendation. Is it also good to read if we study animal microRNAs?
Saurabh Mandal Thanks for your recommendation. However, what I want to know is the method to design primers in software, the tips to know whether primers that we designed is good or bad before making an order in the companies.
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