Drosophila RNAi Screening Center

at Harvard Medical School

Once your screen has been approved, contact Nadire Ramadan (nramadan@genetics.med.harvard.edu) to schedule your screen. (Note that the screening schedule is often full several weeks in advance, so please plan well in advance of the date you would like to start your screen.)

Order of Screening Events at the DRSC:

1. Before arrival to the DRSC, your screen must be optimized in the 384-well format using dsRNA made by the screener (this data must be submitted with your application). The DRSC is unable to provide dsRNA for optimizing screens. Protocols for dsRNA synthesis can be found here
2. Upon arrival to the DRSC, the screener will be trained on the different equipment that will be used during his/her screen.
3. After training, the screener is given one Test Plate to screen. This is to ensure the DRSC staff of the optimization of the screen.
4. Data from the test plate will be reviewed with DRSC staff. If results from the test plate confirm the feasibility of the screen, then and only then may the screener move onto the Pilot screen, which consists of three 384-well plates from our genome-wide collection set. These plates will be screened in duplicate.
5. In order to proceed with the full genome screen, the screener must enter the data from the Pilot screen into the DRSC database. See Ian Flockhart and Matthew Booker for assistance with loading the data. The screener will also meet with DRSC appointed staff to get approval for the continuation of the full genome screen.

Test Plate

During the first week of screening, the first plate the screener will encounter at the DRSC is the Test Plate. This 384 well plate contains a random assortment of 288 genes arrayed in 3 of the 4 quadrants of the plate. The 4th quadrant is left empty for any controls the screener would like to test. This plate will be screened as if it were a part of the full genome screen.

Upon completion of this plate, a meeting will be scheduled with the Director and the Manager to evaluate results from this plate. This meeting will dictate whether or not the screen will be allowed to move onto the Pilot Screen.

Pilot Screen

If the screener has received approval to continue with the screening process, he or she will then conduct the next phase of the screening, which is the Pilot Screen. The Pilot Screen consists of 3 plates out of the Full-Genome set that will be screened in duplicate.

A meeting will once again take place with the screener and the Director and Manager to evaluate the data from this screen. Depending on this result, the screener will be allowed or denied access to the Full Genome Screen.

Genome Wide Screen

If and only if the screen has received approval to continue, the screener will finally be able to move onto the Full Genome Wide Screen. This screen consists of screening the remaining 44 assay plates. Whether the screener performs the screen in duplicate or not is dependent on them.