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Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into...

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Detalles Bibliográficos
Autores principales: McCoy, Adam M., Collins, Michelle L., Ugozzoli, Luis A.
Formato: Texto
Lenguaje:English
Publicado: MyJove Corporation 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2807742/
https://www.ncbi.nlm.nih.gov/pubmed/20057352
http://dx.doi.org/10.3791/1662
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author McCoy, Adam M.
Collins, Michelle L.
Ugozzoli, Luis A.
author_facet McCoy, Adam M.
Collins, Michelle L.
Ugozzoli, Luis A.
author_sort McCoy, Adam M.
collection PubMed
description It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells. We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions. We will demonstrate how to run several samples through a range of electroporation conditions so that an experiment can be conducted at the same time as optimization is performed. We will also show how optimal conditions identified using 96-well electroporation plates can be used with standard electroporation cuvettes, facilitating the switch from electroporation plates to electroporation cuvettes while maintaining the same electroporation efficiency. In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.
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spelling pubmed-28077422012-01-07 Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency McCoy, Adam M. Collins, Michelle L. Ugozzoli, Luis A. J Vis Exp Cellular Biology It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells. We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions. We will demonstrate how to run several samples through a range of electroporation conditions so that an experiment can be conducted at the same time as optimization is performed. We will also show how optimal conditions identified using 96-well electroporation plates can be used with standard electroporation cuvettes, facilitating the switch from electroporation plates to electroporation cuvettes while maintaining the same electroporation efficiency. In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments. MyJove Corporation 2010-01-07 /pmc/articles/PMC2807742/ /pubmed/20057352 http://dx.doi.org/10.3791/1662 Text en Copyright © 2010, Journal of Visualized Experiments http://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Cellular Biology
McCoy, Adam M.
Collins, Michelle L.
Ugozzoli, Luis A.
Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
title Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
title_full Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
title_fullStr Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
title_full_unstemmed Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
title_short Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
title_sort using the gene pulser mxcell electroporation system to transfect primary cells with high efficiency
topic Cellular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2807742/
https://www.ncbi.nlm.nih.gov/pubmed/20057352
http://dx.doi.org/10.3791/1662
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