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transfection of NRK cells-lipofectamine toxicity - (Sep/06/2006 )

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QUOTE (dnafactory @ Sep 7 2006, 02:16 AM)
QUOTE (genehunter-1 @ Sep 6 2006, 11:52 PM)

lotusgirl: Are you trying to tell us different stories in different posts? First, you told us you are not getting good result with IT (10% transfection rate) in this post, however, minutes ago you said IT did great job for you. See below for details in your previous post. In fact you did not even tell the same story at the begining (much lower price) and at the end (cost mounts up to the same level) in this one post. Whats going on?

I have beed regularly using Transit LT-1 from Mirus. It is GREAT! Works much better than Fugene6, at much lower price and less toxixcity to cells. I use NRK cells which are difficult to transfect . Instead of fugene6, I tried Mirus lt-1 I was amazed it worked much better than Fugene6 when I followed their recommended protocol. Mirus usually gives free sample so you cana sk for it and try on your cell line first.

Only drawback. They recommend 7.5 microlitres of the reagent per 35 mm dish per transfection. that means we use more of it per transfection so in the ned the cost mounts up to the same level as of fugene and lipofectamine.

That's not good... Thank you genehunter-1 for finding out...

1. Mirus TransLT-1 is GREAT!!! compared to Fugene 6- it works excellently for immunofluorescence with no toxicity. For IF, 10% efficiency is enough. If you will read my message crefully, it says CLEARLY at the beginning that I need to use Lipofectamine only for western blotting. If u have only 10% efficiency, chances are your transfected protein won't show up on a western blot. But it will be superbright by immunofluorescence.

2. If u are doing only 2 transfections a month, Mirus TransLT-1 is a cheaper option because you have to buy a vial of mirus or fugene 6 even if you are going to use it only once or twice. On the other hand, if u are going to do 500 transfections a mon th ( close to what I do ), Mirus is not so cost-effective because you have to use more of it per transfection, so you end up buying more vials of mirus LT-1 as compared to Fugene6.

Whew!!!! I used to teach 3 yr old kids a long time ago- got the same feeling again, having to explain the economics of reagent buying and the difference between a fluorescence and a biochemical assay!


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