I have run siRNA transfection on SNO cell lines. I seeded in a 6 well plate, incubated at 24 hours and treated with different volumes and concentrations of the siRNA and transfection reagent. I incubated for another 48 hours and extracted. I, then ran cDNA, PCR and a 2% agarose gel but I could not pick up any sign of knockdown, of my target gene as it was expressed as normal.
Does any one have any suggestions to why this is?
It would be appreciated.
Dear Sasha,
48 h is a long time in some cases and ideal for others, so you should collect your samples at different time schedule from 24 to 48 h, then decide the optimum time for your gene.
Another possibility is a technical issue such as the primer you used in PCR, so it is check it again on a negative sample.
- Badges
- Science topic
- Similar topics
- Biological Science
- Physiology
- Physiological Processes
- Stress