Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells

Electrotransformation also known as electroporation is the most reliable and efficient tool for plasmid DNA uptake. Electrotransformation efficiency is function of many factors which include (1) number of cell washes prior to electroporation, (2) electroporation cell number, (3) electroporation DNA...

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Autores principales:	Wu,Ning, Matand,Kanyand, Kebede,Bizuayehu, Acquaah,George, Williams,Sonya
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	cell competency DNA E. coli DH10B electroporation
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500021
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spelling	oai:scielo:S0717-345820100005000212011-05-24Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cellsWu,NingMatand,KanyandKebede,BizuayehuAcquaah,GeorgeWilliams,Sonya cell competency DNA E. coli DH10B electroporation Electrotransformation also known as electroporation is the most reliable and efficient tool for plasmid DNA uptake. Electrotransformation efficiency is function of many factors which include (1) number of cell washes prior to electroporation, (2) electroporation cell number, (3) electroporation DNA amount, and (4) cell growth phase. Those factors have limitedly been concomitantly investigated in E. coli DH10B strain. This study is aimed to explore above key factors to define the optimal conditions for high electrotransformation efficiency. The results showed that electrotransformation efficiency of E. coli DH10B was enhanced to 1.5 x 10(9) cfu/µg by washing cells three times with 15 ml of 10% glycerol. This washed off extra salts from cell suspension and enhanced electrotransformation by preventing arcing and enhancing cell resistance while ensuring minimal level of conductivity. Early exponential phase at 0.15 OD600 was the best growth phase for enhancing electrotransformation of E. coli DH10B. The results also showed that higher electrotransformation efficiency was similarly achieved when 0.5 x 10(10) and 0.6 x 10(10) cell numbers were electroporated with DNA amount ranging from 10 to 40 pg. This study confirmed the optimal conditions for electro competent cell preparation and plasmid DNA electrotransformation, which can result highest transformation efficiency.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.5 20102010-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500021en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cell competency DNA E. coli DH10B electroporation
spellingShingle	cell competency DNA E. coli DH10B electroporation Wu,Ning Matand,Kanyand Kebede,Bizuayehu Acquaah,George Williams,Sonya Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells
description	Electrotransformation also known as electroporation is the most reliable and efficient tool for plasmid DNA uptake. Electrotransformation efficiency is function of many factors which include (1) number of cell washes prior to electroporation, (2) electroporation cell number, (3) electroporation DNA amount, and (4) cell growth phase. Those factors have limitedly been concomitantly investigated in E. coli DH10B strain. This study is aimed to explore above key factors to define the optimal conditions for high electrotransformation efficiency. The results showed that electrotransformation efficiency of E. coli DH10B was enhanced to 1.5 x 10(9) cfu/µg by washing cells three times with 15 ml of 10% glycerol. This washed off extra salts from cell suspension and enhanced electrotransformation by preventing arcing and enhancing cell resistance while ensuring minimal level of conductivity. Early exponential phase at 0.15 OD600 was the best growth phase for enhancing electrotransformation of E. coli DH10B. The results also showed that higher electrotransformation efficiency was similarly achieved when 0.5 x 10(10) and 0.6 x 10(10) cell numbers were electroporated with DNA amount ranging from 10 to 40 pg. This study confirmed the optimal conditions for electro competent cell preparation and plasmid DNA electrotransformation, which can result highest transformation efficiency.
author	Wu,Ning Matand,Kanyand Kebede,Bizuayehu Acquaah,George Williams,Sonya
author_facet	Wu,Ning Matand,Kanyand Kebede,Bizuayehu Acquaah,George Williams,Sonya
author_sort	Wu,Ning
title	Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells
title_short	Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells
title_full	Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells
title_fullStr	Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells
title_full_unstemmed	Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells
title_sort	enhancing dna electrotransformation efficiency in escherichia coli dh10b electrocompetent cells
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500021
work_keys_str_mv	AT wuning enhancingdnaelectrotransformationefficiencyinescherichiacolidh10belectrocompetentcells AT matandkanyand enhancingdnaelectrotransformationefficiencyinescherichiacolidh10belectrocompetentcells AT kebedebizuayehu enhancingdnaelectrotransformationefficiencyinescherichiacolidh10belectrocompetentcells AT acquaahgeorge enhancingdnaelectrotransformationefficiencyinescherichiacolidh10belectrocompetentcells AT williamssonya enhancingdnaelectrotransformationefficiencyinescherichiacolidh10belectrocompetentcells
_version_	1718441825123958784

Enhancing DNA electrotransformation efficiency in Escherichia coli DH10B electrocompetent cells

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