PCR (Polymerase Chain Reaction) is an indispensable technique in the various field of biotechnology. The application of this technique has a great influence on the progress of the bioindustry. In this research, we accumulated the practical and fundamental data for the commercial PCR related products, in order to propose the drafts for international standards of PCR related instrument, reagent and assay method.
Regarding the standardization of the thermalcycler, we tested 4 kinds of thermalcycler concerning the temperature accuracy, temperature uniformity, course of the temperature shift and their effects on the PCR performance. As the result, temperature accuracy and temperature uniformity are supposed to be the items for the standardization.
Regarding the standardization of the reagents, we tested 8 kinds of thermostable DNA polymerase, 3 kinds of reverse transcriptase and 4 kinds of the nucleic acids purification reagent. As the result, sensitivity and fidelity are supposed to be the items for the standardization of thermostable DNA polymerase. Moreover, it is found that the sensitivity and fidelity depend on the type of thermostable DNA polymerase, such as PolI type, alpha type or mixed type. It is appropriate to determine the specification of the sensitivity and fidelity for each type of the thermostable DNA polymerase. Sensitivity and purity of the product are supposed to be the terms to be standardized for reverse transcriptase and nucleic acids purification reagent, respectively.
Regarding the standardization of the assay method for PCR, we tested 2 kinds of nucleic acids detection instrument and 2 kinds of nucleic acids quantification instrument. As the results, sensitivity, dynamic range and reproducibility are supposed to be the terms for the standardization.