Why should primers used in PCR have similar melting temperatures?

Posted June 22, 2020

Answer

Primers in PCR reaction should have similar melting temperatures so that they anneal to and dissociate from complementary DNA sequences at approximately the same temperature, which allows the synchronous extension of both single-stranded DNA molecules in each cycle.

Additional resources

6-ROX glycine *25 uM fluorescence reference solution for PCR reactions*

Chen, S. C., Halliday, C. L., & Meyer, W. (2002). A review of nucleic acid-based diagnostic tests for systemic mycoses with an emphasis on polymerase chain reaction-based assays. Medical mycology, 40(4), 333-357.

Chuang, L. Y., Cheng, Y. H., & Yang, C. H. (2013). Specific primer design for the polymerase chain reaction. Biotechnology letters, 35(10), 1541-1549.