dear all

as it is my first time to design a taqman assay , could you recommend for me a good reference to know the good designing for taqman gene expression assay, what is the accepted range for melting temperature and GC%, and about the second structure of primers.. and how to overcome if i have a problem in the sequence

i used NCBI blast, and primer 3plus to confirm the specificity of the primers, but about the Tm and GC% , i am not sure if they are good to proceed working or not

thanks in advance