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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 18, No. 5, 2015, pp. 1-4
Bioline Code: ej15054
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 18, No. 5, 2015, pp. 1-4

 en A simple and efficient method for extraction of Taq DNA polymerase
Chen, Sique; Zheng, Xiujuan; Cao, Hongrui; Jiang, Linghui; Liu, Fangqian & Sun, Xinli

Abstract

Background: Thermostable DNA polymerase (Taq Pol Ι) from Thermus aquaticus check for this species in other resources has beenwidely used in PCR, which was usually extracted with Pluthero's method. Themethod used ammonium sulfate to precipitate the enzyme, and it saved effort and money but not time. Moreover, we found that 30–40% activity of Taq Pol I was lost at the ammonium sulfate precipitation step, and the product contained a small amount of DNA.
Results: We provided a novel, simplified and low-costmethod to purify the Taq Pol Ι after overproduction of the enzyme in Escherichia coli check for this species in other resources , which used ethanol instead of ammonium sulfate to precipitate the enzyme. The precipitate can be directly dissolved in the storage buffer without dialysis. In addition, DNA and RNA contamination was removed with DNase I and RNase A before precipitation, and the extraction procedure was optimized. Our improvements increase recovery rate and specific activity of the enzyme, and save labor, time, and cost.
Conclusions: Our method uses ethanol, DNase I, and RNase A to purify the Taq Pol Ι, and simplifies the operation, and increases the enzyme recovery rate and quality.

Keywords
Ethanol precipitation; PCR; Purification; Taq DNA polymerase

 
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