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R&D Transfection Insights

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Published on May 18, 2012

Learn more at http://www.lifetechnologies.com/lipof...

Xavier de Mollerat du Jeu, Sr. Staff Scientist at Life Technologies discusses the transfection research program and key factors for successful transfections, which resulted in enhanced protocols for the Lipofectamine® line of transfection reagents.

My name is Xavier deMolleratduJeu, I'm a senior staff scientist at Life Technologies.

It kind of all started with a survey that we collected last year when we wanted to understand what our, the pain point in different microbiology protocols. And when we found out what came out of top analysis, Which was fections, which we thought was pretty routine. But people had a lot of problems with repressability, they were not getting the result they were supposed to get. So this kicked off a new program here that we wanted really to drill down the different factors that affect the transfections. And there's a lot of factors that can affect the transfections. So it took quite a while, but we wanted to understand, you know, what's the importance of cell type on transfections, um, cell passages. Do you really need to optimize to get successful transfections? What's the difference between DNA and sRNA? What buffer you use? All kind of different conditions; all kind of factors, which is incubations, etc. So we wanted to drill this down and really study it. And the goal was to at the end find out, ok, these are the key factors that you need to, to optimize, and you need to, to really pay attention to. But at the end of the day, what we want to do is, people to be the most successful with these transfections. With the least amount of effort. And so that's how we came up with these new protocols, it's much clearer, better colors and really that allows you to do only one experiment and that you will be successful with it.

We kind of simplified, you know, so much clearer, much cleaner formats that really allow anybody to do transfection very easy. But it's not only this. it's not only a cosmetic change with pretty colors. It's really focusing on the key factors for transfection. So when you do this protocol, you should be successful right off the bat.

We've actually done a lot of work. I mean, because there's so many factors that can affect your transfections, from the way you grow your cells to the way you mix to the way you incubate, we really need to understand every factor. So to understand every factor, you really have to test every factor. And the problem, they kind of interact with each other. So really one of the best, the only way to study all those variables at the same time is to do design of experiments in which you can you assess multiple variables at the same time. So we've done all this in the lab, on the wet lab, doing all the transfections and testing all those parameters. And then we plot them out, and we rank each variables in term of importance.

We have this new protocol for three of our main product. One's Lipofectamine 2000. This is the one I just showed you. And the other one is Lipofectamine ltx, which is a new DNA delivery reagents that we recently launched. And the third one is Lipofectamine RNAmax, which is a reagent specifically for sRNA and microRNA delivery.

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