Ready for a Screen?
Think you might be ready to think about a genome wide screen?
(i) Grow up and freeze at least 20+ vials of cells you know work for your assay. These will allow you to go back to standard cell known to 'work' in the future and so be working within the same 'passage range'.
(ii) Scale down your assay into 384 well plates (preferably using the plates specified here - See Plate Types used by facility). Establish the optimum cell number, assay time, and protocol for your assay. Look to maximise the strength of your 'signal’ over the negative controls and look to simplify and
(iii) Contact us directly to purchase assay development plates (See Price List). These 384 well plates are identical to those used for the genome wide libraries, contain dsRNA at the same concentration and purity as the main library.
They will contain 96 negative controls designed to NOT target anything in the Drosophila transcriptome as well as a number of real Drosophila dsRNAs targeting kinases and phosphatases in duplicate. Plenty of spaces for your own assay-specific controls are also left empty. See Plate Description.
(iv) Order / make sufficiently large batches of reagents to allow you to undertake the whole screen (and preferably validation) without mixing batches. For example, grow up sufficient DNA, order batch matched lots of media, serum, antibodies, luciferase substrates, transfection reagents etc. Check everything using assay development plates to be sure they work!
(v) Again, contact us to discuss your preliminary data and quality metrics.
(vi) Fill out and submit the 'Application to Screen' document. This data will be held in strict confidence and is designed to assist the Facility Management Board to schedule screens to schedule screens and be able to ensure that facility time is allocated to genome wide screens with a good chance of success.