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Optimizing Electroporation Parameters to Achieve the Highest Transfection Efficiency

In this series of posts we will be giving tips for developing and improving your own custom electroporation method. Today we will explore Episode 6 - Optimizing Cell Density. Cell density at the time of transfection can significantly affect transfection efficiency. Specifically, if the cells are too dilute or too dense at the time of harvest or the time of transfection, then viability or efficiency may be compromised. Here are some guidelines to help achieve optimal cell densities by cell type. 

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Posted By Michelle M. Ng, Ph. D.

Optimizing Electroporation Parameters to Achieve the Highest Transfection Efficiency

In this series of posts we will be giving tips for developing and optimizing your own custom electroporation method. Today we will explore Episode 7 - Buffer Considerations. The buffer that cells and transfectant are suspended at the time of transfection can affect the viability of cells or the stability of transfectant. Here are some recommendations based on cell type.  

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Posted By Michelle M. Ng, Ph. D.

Optimizing Electroporation Parameters to Achieve the Highest Transfection Efficiency

In this series of posts we are providing tips for developing and improving your own custom electroporation method. Today we will explore Episode 8 - Guidelines For Mammalian Cells. Don’t reinvent the wheel! if there is a protocol or published reference with electroporation parameters for your specific cell line/type (or for a similar cell type, start with it! We offer a Protocol Database searchable by cell type and some General Guidelines for Mammalian Cell Electroporation as well.  

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Posted By Michelle M. Ng, Ph. D.

Optimizing Electroporation Parameters to Achieve the Highest Transfection Efficiency

In this series of posts we will be providing tips for developing and improving your own custom electroporation method. Today we will explore Episode 9 - Guidelines for Bacterial Cells. We offer a protocol for preparing electrocompetent bacteria and some general guidelines for transforming bacteria with electroporation as well.  

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Posted By Michelle M. Ng, Ph. D.


Electroporation Basics

In this post we will explore the relationship between electrical resistance and electrical conductance. We will also provide information on how resistance applies to electroporation applications, and finally we will answer a FAQ on the topic of sample resistance values in electroporation instrument log data. 

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Posted By Michelle M. Ng, Ph. D.


human brainIn this post we will review what organoids are and how they are used to create in vitro models to study organ function and human disease states in three dimensions. We'll cover the topic of organoid transfection via electroporation in more detail. Finally, we will highlight and summarize findings reported by Ogawa et al. 2018. Cell Reports 23, 1220-1229. These researchers utilized electroporation of CRISPR/Cas9 constructs to achieve gene modification of human cerebral organoids, resulting in a glioblastoma tumor model that could be further studied in vivo in xenografted mice.  

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Posted By Michelle M. Ng, Ph. D.


Electrofusion Basics

The joining of membranes of neighboring cells by the application of a pulsed electrical field is called Electrofusion. Electrofusion is widely used for hybridoma creation for monoclonal antibody production, transgenics/somatic cell nuclear transfer applications, hybrid plant creation and functional studies combining two different cell types with different properties.  In this post we’ll explore the types of electrical wave forms used for electrofusion and outline the way that these waveforms are combined to make a successful electrofusion protocol. 

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Posted By Michelle M. Ng, Ph. D.


CAR T-cell therapy

In this post we will highlight a high efficiency method for primary human T cell transfection utilizing electroporation of mRNA.  This method creates genetically engineered T-cells which specifically recognize antigens of interest for CAR T-Cell immunotherapy applications. 

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Posted By Michelle M. Ng, Ph. D.


Electrofusion Basics

There are different ways of analyzing efficiency following an electrofusion experiment. These methods vary in terms of how soon after electrofusion you may analyze results, what types of equipment and reagents are required, and what type of information is determined. In this post we will provide an overview of a couple of analysis methods that provide some initial results just minutes after electrofusion. 

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Posted By Michelle M. Ng, Ph. D.


Utilizing electroporation to brew a better beer

  • In honor of National Beer Day, April 7th, the focus of this blog post is how to use electroporation for genetic engineering of brewing yeast. First, we will provide the ideal electroporation parameters for Saccharomyces cerevisiae using either ECM 630 or Gemini X2 electroporators.  Then, we will highlight an example from the primary literature where this electroporation method was used with brewing yeast Saccharomyces cerevisiae ssp. carlsbergensis to modify the amount of ethanol in beer. 
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Posted By Michelle M. Ng, Ph. D.
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11-20 of 23