Investigating the Effects of DMSO on PCR Fidelity Using a Restriction Digest-Based Method

Abstract

Dimethyl sulfoxide (DMSO) is commonly used in PCR to relieve secondary structures when amplifying high GC templates, but we have little understanding of how it affects PCR fidelity based on its interaction with DNA molecules. To evaluate the possible mutational effects, we developed a restriction digestion-based method to rapidly detect erroneous PCR products and compared our result to normal as well as error-prone PCR conditions. We successfully detected PCR mutations generated by the error-prone condition using three different restriction enzymes. The presence of 2% DMSO in PCR did not appear to affect the yield or the proportion of mutations significantly. Overall, our method is a rapid way to assess the occurrence of mutations in a pool of PCR amplicons and can easily be extended to more sensitive studies.