Analysis of the inhibition of nucleic acid dyes on polymerase chain reaction by capillary electrophoresis

Abstract

An integrated polymerase chain reaction (PCR) and capillary electrophoresis (CE) system can realize accurate quantification of the target PCR product by adding labeling dyes to the PCR reagents, because CE can discriminate all the subsequent nucleic acids, including the primers, non-specific and specific PCR products. Here we discuss the inhibition of labeling dyes on PCR by performing the PCR of Porphyromonas gingivalis (PG) with solutions containing Hoechst 33258, SYBR Green I, and SYBR Green II. Results demonstrated that Hoechst 33258 totally inhibited the PCR process, and PCR efficiency was highly dependent on the concentration of SYBR Green I/II. Such a study expands the capabilities of CE and contributes greatly to the development of hyphenated PCR–CE instruments for biological and medical diagnosis.