A simple and efficient method for extraction of Taq DNA polymerase

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% activ...

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Autores principales:	Chen,Sique, Zheng,Xiujuan, Cao,Hongrui, Jiang,Linghui, Liu,Fangqian, Sun,Xinli
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2015
Materias:	Ethanol precipitation PCR Purification Taq DNA polymerase
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000500005
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spelling	oai:scielo:S0717-345820150005000052016-03-21A simple and efficient method for extraction of Taq DNA polymeraseChen,SiqueZheng,XiujuanCao,HongruiJiang,LinghuiLiu,FangqianSun,Xinli Ethanol precipitation PCR Purification Taq DNA polymerase 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.18 n.5 20152015-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000500005en10.1016/j.ejbt.2015.08.001
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Ethanol precipitation PCR Purification Taq DNA polymerase
spellingShingle	Ethanol precipitation PCR Purification Taq DNA polymerase Chen,Sique Zheng,Xiujuan Cao,Hongrui Jiang,Linghui Liu,Fangqian Sun,Xinli A simple and efficient method for extraction of Taq DNA polymerase
description	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.
author	Chen,Sique Zheng,Xiujuan Cao,Hongrui Jiang,Linghui Liu,Fangqian Sun,Xinli
author_facet	Chen,Sique Zheng,Xiujuan Cao,Hongrui Jiang,Linghui Liu,Fangqian Sun,Xinli
author_sort	Chen,Sique
title	A simple and efficient method for extraction of Taq DNA polymerase
title_short	A simple and efficient method for extraction of Taq DNA polymerase
title_full	A simple and efficient method for extraction of Taq DNA polymerase
title_fullStr	A simple and efficient method for extraction of Taq DNA polymerase
title_full_unstemmed	A simple and efficient method for extraction of Taq DNA polymerase
title_sort	simple and efficient method for extraction of taq dna polymerase
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2015
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000500005
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A simple and efficient method for extraction of Taq DNA polymerase

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