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A simple and efficient method for extraction of Taq DNA polymerase
Chen, Sique; Zheng, Xiujuan; Cao, Hongrui; Jiang, Linghui; Liu, Fangqian; Sun, Xinli.
Afiliação
  • Chen, Sique; Fujian Agricultural & Forestry University. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops. Fuzhou. CN
  • Zheng, Xiujuan; Fujian Agricultural & Forestry University. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops. Fuzhou. CN
  • Cao, Hongrui; Fujian Agricultural & Forestry University. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops. Fuzhou. CN
  • Jiang, Linghui; Fujian Agricultural & Forestry University. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops. Fuzhou. CN
  • Liu, Fangqian; Fujian Agricultural & Forestry University. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops. Fuzhou. CN
  • Sun, Xinli; Fujian Agricultural & Forestry University. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops. Fuzhou. CN
Electron. j. biotechnol ; 18(5): 343-346, Sept. 2015. ilus, graf, tab
Article em En | LILACS | ID: lil-764021
Biblioteca responsável: CL1.1
ABSTRACT
Background Thermostable DNA polymerase (Taq Pol ?) from Thermus aquaticus has been widely used in PCR, which was usually extracted with Pluthero's method. The method 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-cost method to purify the Taq Pol ? after overproduction of the enzyme in Escherichia coli, 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 ?, and simplifies the operation, and increases the enzyme recovery rate and quality.
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Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Taq Polimerase / Etanol Idioma: En Revista: Electron. j. biotechnol Assunto da revista: BIOTECNOLOGIA Ano de publicação: 2015 Tipo de documento: Article País de afiliação: China

Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Taq Polimerase / Etanol Idioma: En Revista: Electron. j. biotechnol Assunto da revista: BIOTECNOLOGIA Ano de publicação: 2015 Tipo de documento: Article País de afiliação: China