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siRNA
Posted by: Sushi (IP Hidden, New member, 1)
Date: July 1, 2005 07:17PM
hi,
I will transiently transfect my cell with siRNA oligo. I plan to dected the expression of target gene and its protein 1-5 days post transfection. Because cell will devide, will this lead to the more cells do not contain siRNA? If so, do I need transfect siRNA everyday during 5-day analysis.
Re: siRNA
Posted by: thomas (IP Hidden, New member, 5)
Date: July 3, 2005 02:16PM
yes the siRNA effect is transient and when cell divides it splits siRNA between two cytoplasms
also notice that for most proteins - you can detect protein knockdown only 3-4 days after transfection
Re: siRNA
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: July 7, 2005 11:27AM
by 5 days the protein level should have returned to normal (before siRNA transfection). whether to transfect everyday depends on the goal of your experiment. do you wish to measure the stability of transfection over the course of 1-5 post transfection? how the level of protein changes postransfection? how long long the silencing lasts? if yes to either of these questions: do not transfect everyday, as you will not be able to determine the answer.
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