Experimental Workflow:
Using siRNAs
to Delineate Gene Function
Step 1. Obtain effective siRNAs
To obtain gene silencing, potent and specific
siRNAs must be designed. Effective siRNA design and rigorous
quality control of synthesis and purification will prevent the waste of both time and money.
Efficient, reproducible siRNA
delivery is essential for successful RNAi experiments.
Because siRNA delivery is so critical, it is highly
worthwhile to invest the time and effort to determine
the best siRNA delivery method and conditions for
each cell line studied.
Once siRNAs are obtained and delivery is optimized, the next step in
a typical gene silencing experiment is to assess the silencing
efficiency of the selected siRNAs.
Step
5. Correlate silencing efficiency with biological effect
In order to fully understand the biological
impact of reduced target gene expression, it is necessary
to correlate the observed phenotype with the level of knockdown
induced.
