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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...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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MyJove Corporation
2010
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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. |
format | Text |
id | pubmed-2807742 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
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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