cDNA Library Screening
Our Laboratory Automation group is now providing a high-throughput screening service utilizing a new plasmid-based, genome-wide human cDNA library. This new expression library was created in an extensive collaboration between Dr. Brian Seed's Laboratory and the CCIB DNA Core Team. Our current library version consists of 19,096 sequence-verified, full-length cDNA clones but we are still adding new clones.
If you are interested in performing a screening project in collaboration with the CCIB DNA Core, please contact us.
-------------------------------------------------------------------------------------------------------------------------------
Screening Process:Our screening process is designed to be a highly collaborative endeavor between investigator and core personnel (user-assisted model).
Responsibilities of the investigator include:
- Development and validation of screening assay in 96-well format (incl. reporter construction, positive and negative controls)
- Plating of cultured mammalian cells (in 96-well format) throughout the entire screening porcess (to be performed at researcher's home institution)
- Providing screening assay reagents (Luciferase-based assay system preferred), reporter constructs and controls
Responsibilities of the CCIB DNA Core include:
- Pilot screen to confirm feasibility
Should the pilot screen yield unsatisfactory results, the researcher will engage in further assay optimization at his/her home institution. - Completion of screening project, including data analysis and top hit identification
To measure gene expression levels (i.e. gene activation or inactivation), luminescence read-outs will be performed using the TopCount NXT plate reader. The identities of the top hit clones are determined by statistical analysis.