High Throughput Screening

Description

https://www.ncl.ac.uk/htsf/

The High Throughput Screening Facility in the medical faculty at Newcastle University possesses a range of robotic equipment for high throughput biological screens and can assist with experimental design and statistical analysis.

Services Offered

■siRNA and mammalian screening: siRNAs can be cherry picked from libraries. siRNAs can be transfected into mammalian cell cultures.

■Synthetic Genetic Array (SGA): A query gene mutation(s) is combined with all non-essential null mutations in the budding yeast genome to produce 5000 different double mutant yeast strains. Fitness of all double mutants is determined by photography.

■Quantitative Fitness Analysis (QFA): Determines fitness of thousands of parallel microbial cultures growing on solid agar plates. QFA can be used to assess the effects of genetic, drug, temperature or nutrient interventions. Typically we quantify cell density of 384 independent cultures per plate and determine the exponential-phase growth rate, saturated culture density and fitness for all strains.

■Libraries and Culture Handling: We have single-deletion and GAL overexpression libraries from S. cerevisiae, and we are happy to discuss the purchase of other libraries. We can also handle many other biological materials (e.g. siRNA, bacterial, S. pombe or mammalian cells).

Libraries Retained

■Dharmacon Human Drugable Genome Library

■GPCR

■Protein Kinases

■Ion Channels

■Phosphatases

■Drug Targets- Includes genes involved with apoptosis, senesence, nucleic acid binding, autophagy, DNA repair, and characterized nuclear receptors

■Proteases

■Ubiquitin Conjugation 1 - Cullins, E1, E2, HECT E3 Ligases

■Ubiquitin Conjugation 2 - F-box, SOCS box E3 Ligases

■Ubiquitin Conjugation 3 - RING finger and RING finger-like E3 Ligases

■Synthetic Genetic Array (SGA): A query gene mutation(s) is combined with all non-essential null mutations in the budding yeast genome to produce 5000 different double mutant yeast strains. Fitness of all double mutants is determined by photography.

■Quantitative Fitness Analysis (QFA): Determines fitness of thousands of parallel microbial cultures growing on solid agar plates. QFA can be used to assess the effects of genetic, drug, temperature or nutrient interventions. Typically we quantify cell density of 384 independent cultures per plate and determine the exponential-phase growth rate, saturated culture density and fitness for all strains.

■Libraries and Culture Handling: We have single-deletion and GAL overexpression libraries from S. cerevisiae, and we are happy to discuss the purchase of other libraries. We can also handle many other biological materials (e.g. siRNA, bacterial, S. pombe or mammalian cells).

Equipment

■Beckman Biomek FX Liquid Handler: Pipettes and mixes liquids and cultures. Consists of 96 well and Span-8 (1-8 independent channel) liquid handling tips. Can transfer a single or up to 96 liquids between media or onto solid agar. The FX is integrated with a stacker allowing use of up to 189 plates per run, vortexer, bar code readers and tip washing stations.

■Biomatrix BM3-09 Colony Handler: Transfers solid microbial colonies in 1, 96, 384 or 1536 colony formats from solid agar to solid agar or to liquid culture (e.g. 96 well plates). The BM3 can replicate or re-array libraries, revive frozen libraries and an integrated stacker up to 192 plates per run and contains an integrated bar code reader.

■Matrix WellMate Dispenser: For rapid, sterile dispensing of liquid and cultures in 96 or 384 well format. Handles volumes from 1μl to 2000μl.

■spImager: High resolution agar plate imaging system with uniform illumination, reproducible plate positioning and integrated bar code reader.

■S&P Robotics Automated Incubators: Automated agar plate photographic system. Allowing regular periodic (e.g. every 4 hours) photography of up to 160 agar plates for periods up to one week. Contains integrated bar code reader.

Contact: peter.banks@ncl.ac.uk