Pilot production of Taq and Pfu DNA polymerases

Document Type : Research Paper

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Abstract

Taq and Pfu DNA polymerases are two important enzymes in molecular biology studies. They are widely used in all molecular biology laboratories. Simple method of production and purification of the two enzymes is reported here to locally supply laboratories with high consumption level. Plasmids containing the Taq and Pfu genes were separately transformed to an E. coli strain. The transformed bacteria were grown on liquid 2x SOB culture medium for 4.5 hours. Bacterial cultures were induced to produce the enzymes. Extraction and purification of the enzymes was conducted as presented in this study. Bradford assay was used to determine the enzyme concentration. The activity and efficiency of Taq and Pfu enzymes produced here were compared to a commercial enzyme by amplifying a known genomic fragment of Arabidopsis thaliana ecotype Colombia. Therefore, the enzyme production method adopted in this study was efficient in producing high purity Taq and Pfu

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