Why should I worry about, and how do I minimize primer dimer formation, in my PCR assays?
Primer dimers are undesirable side-products of PCR and result when one primer anneals to another primer, forming a substrate for amplification by the DNA polymerase during PCR. This secondary product contributes to the SYBR Green qPCR signal for a gene, artificially increasing its apparent expression level. In end-point PCR, the amplification of the primer dimer can decrease the level of PCR reagents needed for the amplification of the gene-specific product of interest, thereby artificially decreasing its apparent expression level. Rigorous optimization of qPCR Primer Assays and qPCR master mixes can lead to the development of optimal assay conditions, which virtually eliminate the production of primer dimers.
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- Why should I worry about, and how do I minimize primer dimer formation, in my PCR assays?
