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RNAi and SiRNA Forum Discuss and Post Questions in this forum about RNA interference and siRNA. RNAi and siRNA transfection, inhibition of endogenous genes, and miRNA and shRNA.


Problem with siRNA transfection

Problem with siRNA transfection - RNAi and SiRNA Forum

Problem with siRNA transfection - Discuss and Post Questions in this forum about RNA interference and siRNA. RNAi and siRNA transfection, inhibition of endogenous genes, and miRNA and shRNA.


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  #1  
Old 03-29-2008, 08:37 PM
Pipette Filler
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Default Problem with siRNA transfection



Hello All,
Actually ,since three month ago I have tried to transfection siRNA into HUVEC and HASMC cells but I could not get any significant reduction in mRNA .I used Lipofectamin 2000 from invitrogen.Could anyone know or has experience with these cells.By the way I used 6 well plate and the amounts of Lipofectamin were 1, 2, 2.5 and 5ul and siRNA concentration was 20um (25ul and 50ul).
Hope I will hear the solution.
Ela
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Old 04-02-2008, 11:49 AM
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Default Re: Problem with siRNA transfection

Dear Ela,

All siRNAs are not equally efficient. How many have you tried ? You can flourescently label the siRNA and then transfect. Later you can check for its entry by microscopy.

Moreover you can try using siPORTamine from ambion which is optimized for delivery into primary cells.

Regards,
Sumit
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  #3  
Old 04-03-2008, 09:47 AM
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Default Re: Problem with siRNA transfection

Hello ,
Thanks , I used ,1ul ,2ul ,2.5ul 3ul , 5ul,15,25 and 37.5ul,the conc of my siRNA is 20 um.
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