Hello, I am working on microRNA expression studies on Regeneration. I isolated RNA from my sample and converted it into cDNA using Poly A polymerase and Reverse transcriptase enzyme. Previously I designed a miRNA-specific Forward primer at the melting temperature of 60°C. For Reverse primer, I used Universal Reverse primer from a commercial kit. But now I need to design a miRNA reverse primer for myself. Kindly suggest me method to design a reverse primer for Poly A-tailed miRNA. Thanks in advance.

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