How important is considering the exon-exon junction for primer design in order to perform RT-real time PCR by SYBR-green?

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Laurence Stuart Dawkins-Hall

Very important
By designing exon junction primers cDNA and not genomic DNA is specifically targeted
Primers can be situated internally within exons but you have to ensure all genomic DNA is eliminated by DNAse I digestion

Kunnathur Murugesan Sakthivel

Thank you for your valuable comments Dr,Laurence Stuart Dawkins-Hall and Dr.Maureen J Ostaff and