Typically siRNA screening programs are run using pooled sets of at least three siRNAs that are supplied in either a 96 or 384 well plate format and screened in a variety of ways. Hits are identified and the individual siRNAs from that pool
are then re-transfected as single siRNAs and tested again to determine which siRNA elicited the effect.
This approach faces several technical challenges:
- arrayed siRNA libraries and transfection reagents are costly
- libraries need to be reformatted from 96 well source plates to 384 or 1,536 well screening plates
- the generation of phenotypic data is slow and requires significant infrastructure investment (automated microscopes, data processing cores, dedicated data storage servers)
Finally, the process of identifying and selecting the individual hit siRNAs selected from the screen, typically from a bank of -80°C freezers has two main drawbacks:
- very labour intensive process
- runs the risk of damaging the integrity of the sample due to warming when the freezer doors are opened and the tubes located, RNAs are prone to temperature-induced degradation so this is a big concern
Here we present an integrated solution from TTP Labtech which overcomes these challenges and enables robust and fast genome-wide phenotypic screens.