I'm looking for exact information even like si-RNA sequences and how to work on it.
IκB is the inhibitor of NF-κB. Why would you like to turn off both of them? There are studies for turning off NF-κB by siRNA (http://www.sciencedirect.com/science/article/pii/S0166093404001260), but to include two siRNA into medium and being sure that every cell has copies of each siRNA for required amount is very hard.
I'm not ganna turn off both of them. actually it was I-kB kinase. I improved it. thanks for your help. I'm waiting for your offers.
One other way is to use the first siRNA and then after the first passage of the cells you can add the second siRNA. By the third passage you have to have both of mRNAs turned down. The results depends on many different factors, including the specificity of siRNAs you are using and also the cell type. Good Luck.
Hi mitra
these siRNAs are toxic, I can't wait for that. that's my big problem
May be you need to optimize your transfection protocol to lower the toxicity. I have worked with siRNA before, and I didn't see that much toxicity at least during the first week after transfection. Sometimes by using a lower amount you can keep the cells for a longer time while waiting for the siRNA to work.
I suggest you search the literature for different protocols first and see which one works better for your cell line. you may spend some time in the beginning to optimize your protocol, but at the end it will be worthwhile. I have attached a few protocols here for you to look at. Also, below are some links you can use.
http://www.lifetechnologies.com/us/en/home/life-science/cell-culture/transfection/rnai-transfection/rnai-transfection-protocols.html
http://www.sigmaaldrich.com/life-science/functional-genomics-and-rnai/sirna/learning-center/mission-sup-reg0/experimental-design0.html
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