A simpli�ed protocol for producing Taq DNA polymerase in biology laboratory

Farazmandfar, Touraj and Rafiei, Alireza and Hashemi-Sotehoh, Mohammad Bagher and Valadan, Reza and Alavi, Mohammad and Moradian, Fatemeh (2013) A simpli�ed protocol for producing Taq DNA polymerase in biology laboratory. Research in Molecular Medicine, 1 (2). pp. 23-26.

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Official URL: http://rmm.mazums.ac.ir/article-1-40-en.html

Abstract

Background: Taq DNA polymerase is a very important enzyme for molecular biological studies such as DNA amplification and DNA sequencing by the PCR. It is a standard enzyme that is used in 90 of molecular biology labs today. The aim of this study was to produce Taq DNA polymerase enzyme in E. coli by a reliable, practical, simple and low cost method. Materials and Methods: In this study, the Taq gene was amplified from the genomic DNA of Thermus aquaticus and cloned into pTrc99A vector. Recombinant plasmid is expressed in E. coli strain TOP10. Product protein is extracted and purified. Expression of gene was analyzed by SDS-PAGE and gene amplification. Results: SDS-PAGE showed an approximately 94 KDa protein. The density of protein bands in agarose gel electrophoresis indicated that the purified enzyme is more active than the non purified one. Conclusion: The protocols described in this paper lead to the production of pure and active enzyme that can be applied in both teaching and research laboratories.

Item Type: Article
Uncontrolled Keywords: Taq polymerase, expression, Purification
Depositing User: Unnamed user with email eprints@mazums.ac.ir
Date Deposited: 04 Jan 2018 14:18
Last Modified: 04 Jan 2018 14:18
URI: http://eprints.mazums.ac.ir/id/eprint/1570

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