I want to check the role of one protein in neuronal cell line. For this, I'm going to use siRNA knock down of gene for that protein, in order to check whether knock down gene can effect on cells or not.
Can anyone give me the brief detail of the procedure of siRNA interference and the protocol from the first step, how treat cells with siRNA and lentivirus, incubation time, how to calculate the amount of materials (viral particles and others) that should add to the cell cultures, number of controls, and how to harvest cells?
Thanks in advance.
If transfection is well standardized in your cell line, go for transfection reagent based siRNA knockdown. 72 hours treatment is enough for knocking down.
Dear Rupam, Thanks for your reply. Actually, I would like to use the MISSION lentiviral transduction particles to siRNA treatment in neuronal cells. I'm looking for protocol to use. I would like to know what I have to do before process and during the process.
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