Plate Description

The plates we use at the SRSF are SBS format, tissue culture ready, sterile with an excellent signal-to-noise ratio. All assay, subset-library and library plates come with 5ml of dsRNA, dispensed into the bottom of each well. White plates are used for luminescent detection, black for fluorescent plate reader detection and black-clear bottomed for microscopy acquisition. Each holds approximately 120ml of liquid, although we prefer not to use more than 100ml if we are using disposable seals from the heat-sealing plate machine (Agilent PlateLoc).

Each of the plate types we supply has the generic SRSF format (see figure). That is, each well contains ~250ng of dsRNA except the wells used for control purposes and the 4 top left-hand corner wells, which are dsRNA-free controls. The 16 positive and negative control wells are ready for users to spike in their own controls. ie, dsRNAs targeting genes that may or may not have an affect upon the process that you are investigating.  The Library and sub-set plates only use these wells as the source of controls, but the “assay development plate” has 94 internal controls (mainly negative), so we suggest for users to test more positive controls in these 16 “spike-ready” wells (they are the blacken wells in this figure).

Another aspect of our plates is that they contain 4 wells, which position moves between plates, of dsRNA targeting the Drosophila inhibitor of apoptosis gene, DIAP1. When DIAP1 is knocked down, cells initiate the apoptotic cells death program. DIAP1 makes an excellent visual bar-code and dsRNA control, so that we can check plates have not been mixed-up and dsRNAs are functional. This enables us to determine the content of each plate even when the typed-barcodes are accidentally lost from each plate.

Generic Plate