Different siRNA transfection agents - Have enyone tested them??? (Dec/16/2004 )

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QUOTE (kevin77 @ Aug 17 2005, 09:49 AM)

I have worked using siRNA with Jurkat cells and K562 cells in my previous lab. For such suspension cells, normally lipid-based transfection reagents have presented very low transfection effiencies. I have tested OpenBiosystem Arrest-In and Roche x-tremeQ2 (for K562, Jurkat and HeLa), both got <10% transfection efficiency, and Arrest-In is more toxic.
For that reason, we turned to eletroporation-Amaxa Nucleofector, after optimization, for early passage Jurkat cells (<10), we could get >90% efficiency.
Good Luck!
kevin

How much viability were you getting after electroporation? Are you doing transient or stable transfection? I'm starting work with Jurkat cells with the hope of transiently transfecting the cells. If I can stably transfect them, even better. Therefore any wisdom is appreciated! Thanks.

-kaustav-

Has anyone tried siRNA on mouse embryonic stem cells?

-PhDStudentSweden-

Hi,

I am having problems for transfecting my 3T3 L1 cells... anyone got any positive results with these cells??

Thanks..

Suman

-sumandas-

Has anyone had any experience with the human myeloma cell line RPMI 8226? I was working on transfecting them with RNAifect and am getting a pretty severe growth inhibition. I am planning on testing different transfection media but would like to have a better idea of where to start. If anyone has worked with this, or a similar cell line I would love to hear if you have had any success.

-jphipps-

QUOTE (jphipps @ Nov 5 2007, 08:32 AM)

I maybe wrong on this, but I dont think most transfection reagents work well for suspension cells. Electroporation or lenti viral would be a better choice.

-genehunter-1-

QUOTE (genehunter-1 @ Nov 5 2007, 08:47 AM)

QUOTE (jphipps @ Nov 5 2007, 08:32 AM)

I maybe wrong on this, but I dont think most transfection reagents work well for suspension cells. Electroporation or lenti viral would be a better choice.

That is true to some extent, although the protocols for most reagents now support suspension cells. Besides, the issue isn't transfection efficiency but cytotoxicity.

-jphipps-

oligofectamine is good for Hela cells. Liptofectamine 2000 may need to change the medium after transfection otherwise it is toxic to Hela.

-tulip1-

Hi there.
I'm performing silencing experiments using siRNAs and I will have to transfect SK-N-BE neuroblastoma cell line.
I've experienced very low transfection efficiency and a moderate toxicity using QIAGEN Hiperfect. I'm going to try lipofectamine and metafectene too.

Now here's my question: is there anyone of You that has experience in transfecting SK-N-BE cell line? Any experience with lipofectamine and metafectene, by chance?
Any hint would be very appreciated.

Thanks in advance.

-HP1-

Anyone did siRNA transfection on primary fetal mouse hepatocytes?

I am trying siPORT lipid and no fluorescence is observed....

Any suggestions?

Like switching to Lipofectamine 2000?

-jiro_killua-

Hi,
I am new in the field of siRNA, and I was wondering if you have any protocol to transfect HEK293 cells with siRNA using the calcium phosphate method. Thank you

Patricio

-patricio-

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