GB/T 35541-2017 in English

Interpretation of the National Standard of the People's Republic of China GB/T 35541—2017

Definition and Characteristics of pfu DNA Polymerase

pfu DNA polymerase is derived from Pyrococcus firosus and has extremely high thermal stability and fidelity. It can catalyze the polymerization of deoxyribonucleotides from the $5^{\prime}{-3}^{\prime}$ direction of the template to the primer, has $3^{^{\prime}}{-}5^{^{\prime}}$ exonuclease activity, and has no $5^{\prime}{-}3^{\prime}$ exonuclease activity. Its molecular weight is about $90~\mathrm{ku}$.

Background of Standard Formulation

With the rapid development of genetic engineering and biotechnology, pfu DNA polymerase is increasingly used in molecular biology experiments. This standard is formulated to regulate the production and use of this type of tool enzyme and promote its industrial development.

Interpretation of Technical Requirements

Test methods and experimental steps

Comparison of standard framework and analysis of technological evolution

Compared with previous standards, GB/T 35541-2017 has achieved important breakthroughs in the following aspects:

• More stringent enzyme activity detection methods have been introduced;
• The impurity detection process has been optimized, especially the detection of exonucleases and endonucleases;
• Specific requirements for reagent preparation and experimental conditions have been added.

Implementation recommendations

To ensure the effective implementation of the standard, it is recommended that:

1. Manufacturers should strengthen internal training to ensure that technical personnel are familiar with the requirements of the new standard;
2. Testing institutions need to be equipped with advanced PCR analysis systems and related reagents;
3. During the production and storage process, operations should be strictly carried out in accordance with freezing conditions to avoid enzyme activity loss.