High-throughput Screening

The High-throughput Screening (HTS) facility provides a core service for LRI research groups. It enables individual research groups to use large-scale screening technologies and approaches as part of their research arsenal. It is staffed by highly experienced researchers who collaborate closely with LRI research groups in developing assays for screening. The facility currently specialises in genome-wide siRNA and RNAi screens in mammalian and Drosophila cells and has both a human siRNA Smartpool genome library and a corresponding deconvoluted library from Dharmacon, as well as a Drosophila genome RNAi library from Ambion. In addition the service offers researchers the opportunity to screen a range of well-characterised chemical inhibitors from both the LOPAC1280 collection (Sigma) and kinase inhibitor collection (Calbiochem).

Once an assay is sufficiently robust for a screening campaign, the facility¿s staff conduct the screen and collect and analyse the data. To help them do this, the facility has a number of high-end machines enabling researchers to utilise a wide range of assay formats and experimental designs. For example, liquid handling is automated using:

• Dual 96-well head Biomek FP liquid handling robot (Beckman) in a Class II cabinet (Bigneat) and linked to a Cytomat 24 incubator and Cytomat hotel (ThermoFisher).
• BioTek ELX405 plate washer (BioTek)
• Wellmate liquid dispensers (ThermoFisher)

Data acquisition is automated using:

• PheraStar plate reader (BMG Labtech)
• Acumen Explorer eX3 laser scanning microplate cytometer (TTPLabtech)
• ArrayScan automated microscope (Cellomics) with on-the-fly image analysis capability

Data analysis utilizes a number of in-house data management and analysis solutions and freely available Bioconductor analysis packages.

As part of its remit to undertake any type of large-scale screening activity that might be of benefit to LRI research groups, the facility has recently begun offering groups the chance to perform synthetic lethal screens in both S. cerevisiae and S. pombe using collections of viable deletions in both organisms and a QPixXT colony picking and replicating robot (Genetix).