Amaxa

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Paul
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Amaxa

Does anyone have any experience using the Amaxa electroporator? What cells/ % transfection/level of expression?
Paul

mijst4
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mijst4 wrote:Paul wrote: Does

mijst4 wrote:

Paul wrote:
Does anyone have any experience using the Amaxa electroporator? What cells/ % transfection/level of expression?
Paul

We have just received one for demo, and we trasfected GFP to K562 cells. The transfection efficiency with typical transfection methods was usually less than 5%, but with amexa electorporator, it was over 60%, and the cells look really well.

got777
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the amaxa electroporator is a

the amaxa electroporator is a very good machine. What they say on their web site seems to be right
the main problem is the price 11000 $ and around 300$ for 25 transfection (1 million of cells each)

samm
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Am about to try out a rather

Am about to try out a rather difficult transfection (stable, 19Kb) - if that works, the AMAXA has my wholehearted unreserved support! Sadly, most of the "sample" sets around are with the pMAX GFP supplied by AMAXA. What kind of efficiencies are people getting with other vectors, different cell lines, and co-transfections? Perhaps there should be a new topic devoted to researchers experience with this new technology. moderators - help!
p.s. just sent out the order for AMAXA - worked out 336USD (incl shipping in US) for oone kit (25 transfections)

Fraser Moss
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I have the device in trial

I have the device in trial right now

I can get 90% transfection efficiency of the ATCC CHO-K1 cell line with the pmaxGFP plasmid. (1, 1.5, or 2 micrograms in 1 million cells)

I have also transfected a hERG K+ channel and got at least 60% efficiency.

Basicaly, if you are transfecting cDNA it is excellent.

I have also found that you can actually scale down the transfection volume to 50ul from the reccomended 100ul (if needs be), especially if your application does not need all the cells in the dish to be transfected (e.g. patch clamp electrophys), in which case you can plate your transfected cells into dishes that already contain about 250000-300000 nontransfected feeder cells.

jonatmudd
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I've tried AMAXA with E18 rat

I've tried AMAXA with E18 rat hippocampal cells to transfect GFP (AMAXA's "pmax GfP").
Their website claims 40% transfection efficiency. I got no where that close in 4 different trials on 4 different weeks. I was getting more like 15%.

AMAXA is annoying because they keep all of their protocols proprietary. When I called for help all they could offer were cryptic other programs like "O-12 and T-7." Couldn't even try to optimize myself.

Definitely not recommended for primary brain cells.

Tony Rook
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I've heard from several

I've heard from several source besides this thread that AMAXA electroporator's have much better efficiency in regards to transfection/transformation. Can anyone explain what make their system work so much better than other electroporators on the market?

samm
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Hi Tony! AMAXA uses a

Hi Tony! AMAXA uses a proprietary shaped pulse charge, coupled with their buffered carriers, to get DNA complexes straight into the nucleus. This works spectacularly well for some genes (e.g. GFP, b-gal) in difficult-to-transfect cells like MDCK (60-85% routine) - however, does not work well for some other genes - esp large ones. Also, you have to play around to optimize the protocol (code) used for your cells/specific gene.