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?
Sv Mg
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..
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