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transfection using lipofectamine 2000


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#1 luckyjade

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Posted 21 February 2014 - 12:22 AM

Hi,

 

I have a question about the use of lipofectamine 2000 for transfection of 293t cells. i am performing the transfection to confirm for functionality of certain plasmids. i am wondering if the amount of lipo 2000 that i put into a well makes a significant difference? the protocol recommends 1 uL-2.5 uL of lipo for each well and i only used 1 uL. will this make a significant difference? what effects are there for using a smaller or greater amount of lipo?



#2 luckyjade

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Posted 21 February 2014 - 12:23 AM

*this is for 24 well transfection



#3 luckyjade

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Posted 24 February 2014 - 06:33 PM

forgot to add: lipofectamine 2000 is from invitrogen



#4 bob1

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Posted 25 February 2014 - 02:57 PM

Usually it is the ratio of lipofectamine:DNA that is the important thing.  Most protocols call for a 3:1 ratio (ul lipo:ug DNA), but different ratios should be tried for each plasmid to optimize the transfection. 

 

However, these reagents are toxic to the cells, so the more you use, the more toxicity you will see.






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