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Mission Impossible: Getting Into the Cell Alive

January 8, 2018 11:17:04 AM EST

Getting Into the Cell

Transfection is simply getting DNA (or other molecules of interest) into a cell, while keeping that cell alive. We can do this in countless methods which are divided into three main categories; Reagent Based, Instrument Based, Biological Based. Each method has its own advantages and disadvantages and may be used for transient or stable transfections. Read on as we indulge ourselves into the top methods to see which one suits you best.

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Posted By Michelle Ng, Ph. D. & Chelsea Nelson

Catching the Electroporation Wave

September 25, 2018 2:43:53 PM EDT

Getting Into the Cell

Electroporators can come in multiple electrical waveform styles and typically allow you to vary the characteristics of the electrical pulse settings. Every cell type is unique in terms of which pulse characteristics work best. Field Strength (€, usually expressed in V/cm) is dependent on the pulse parameters applied (voltage, capacitance and resistance) and the distance between the electrode or cuvette contacts. Application of this electrical field causes Electropermeabilization (transient pores in the cell membrane through induction of transmembrane voltage) allowing nucleic acids to pass through the cell membrane.

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Posted By Michelle M. Ng, Ph. D.

Targeting Invaders Using CAR-T Cells

October 26, 2018 3:32:00 PM EDT


Creating CAR-T Cells with Electroporation

The Immune system is the body’s defense against infections and cancer. It consists of billions of cells that are divided into several different types. A subtype of white blood cells calls Lymphocytes comprise a major portion of the immune system. There are three types: B lymphocytes, T lymphocytes and Natural Killer (NK). In today’s blog post we focus on T lymphocytes (T cells) and how they work to kill tumors. 

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Posted By Michelle M. Ng, Ph. D.

Unleash the Power of CRISPR with Electroporation

March 12, 2019 4:52:00 PM EDT

CRISPR for Gene editing knockout diagram

The CRISPR/Cas system is a prokaryotic immune system that allows the cell to protect itself from foreign DNA such as a virus or plasmid. Modified versions of CRISPR/Cas9 are being utilized as gene editing tools in revolutionary ways in science and medicine.

Here we explore the formats that CRISPR/Cas9 tools and the benefits of using electroporation to deliver CRISPR gene editing constructs. 

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Posted By Michelle M. Ng, Ph. D.

Improving Cell Viability During Transfection

March 28, 2019 4:52:00 PM EDT

Improving Cell Viability During Transfection

There are several common causes of high cellular toxicity following electroporation.  Here’s a list of the top causes of cell death during electroporation and recommended solutions.  

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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 provide tips for developing or improving your own custom electroporation method. Today we will explore Episode 1 - Converting between Square and Exponential Decay Waves.  

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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 or improving your own custom electroporation method. Today we will explore Episode 2 - Converting Between Different Gap Sizes. Today we will explain how to convert between two electrodes or cuvettes with different gap sizes.  

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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 or improving your own custom electroporation method. Today we will explore Episode 3 - Scaling Up and Down. There is some room for scaling electroporation volumes up and down just by switching cuvette gap sizes. For even larger volumes you will need to move to a different type of chamber.  

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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 or improving your own custom electroporation method. Today we will explore Episode 4 - Temperature Considerations.The majority of mammalian cells are electroporated efficiently at room temperature. However there are couple reasons to consider an electroporation experiment at a lower temperature such as 4 ͦC.  

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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 or improving your own custom electroporation method. Today we will explore Episode 5 - Considering Transfectant Amount. Transfectant is the molecule of interest that you wish to deliver to your target cells or tissues. The amount of transfectant used in your experiment can impact transfection efficiency. Increasing the amount of transfectant can boost efficiency up to a point; determining the optimal amount to use by running a titration experiment can be helpful.  

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Posted By Michelle M. Ng, Ph. D.
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