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Using the gene pulser MXcell electroporation system to transfect primary cells with high efficiency.

McCoy AM, Collins ML, Ugozzoli LA - J Vis Exp (2010)

Bottom Line: It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells.We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions.In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.

View Article: PubMed Central - PubMed

Affiliation: Gene Expression Division, Bio-Rad Laboratories, Inc.

ABSTRACT
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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Using the gene pulser MXcell electroporation system to transfect primary cells with high efficiency.

McCoy AM, Collins ML, Ugozzoli LA - J Vis Exp (2010)

© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2807742&req=5

Bottom Line: It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells.We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions.In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.

View Article: PubMed Central - PubMed

Affiliation: Gene Expression Division, Bio-Rad Laboratories, Inc.

ABSTRACT
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.

Show MeSH