High-Throughput Bioscience Center (HTBC) Fully automated integrated system: –8, 96 or 384 tip pipettors/pin tools (CaliperLS SciClone ALH 3000) –Robotic arms/Stackers for moving microplates (Zymark Twister II) –Robotic 198 plate CO 2 Incubator –Integrated plate washer/reagent dispenser (Bio-tek EL406) and reagent dispenser (Multidrop) –Bar Code reader –Microplate reader (Molecular Devices AnalystGT, Tecan Infinite M1000) Access to rapidly test thousands of molecules in various bioassays. –Small Molecule Compound library—114,000 diverse compounds (ChemDiv, Chembridge, Specs, ChemDiv Kinase) –Drug Repurposing Libraries—8,770 compounds of known modulators/FDA approved Drugs (LOPAC, NIHCC, NCI DTP, Micorsource, Biomol ICCB and FDA) –Small interfering RNAs (siRNAs)—Whole genomes targeting 21,000 genes, pools of 4 siRNAs/gene, Human library from Dharmacon, and Mouse library from Qiagen –cDNAs—15,000 genes, E. coli stocks Human ORFeome in Gateway vectors from Open Biosystems High-Throughput Assays (384-well plates) –In vitro targets--Enzymes, Protein-protein interactions, Ligand-receptor binding, Protein- DNA binding, etc. (10-15,000 compounds/day) –In vivo cell based-spectrophotometric-- Reporter gene-luciferase, proliferation, synthetic lethality, etc. (15-30,000 compounds per week) –High-Content Imaging--Mitosis, protein translocation/location/expression, cellular morphology, apoptosis, (5-10,000 compounds per week)

siRNA Screening at the HTBC siARRAY whole Human Genome siRNA library from ThermoFisher (formerly Dharmacon) –80 gene targets per plate 4 siRNA oligos per gene--pooled in each well 267 X 96 well plates targeting 21,122 genes –Library subdivided in: 10 Kinase (779 genes, can be tested separately) 7 GPCR (517 genes) 76 Druggable genome (6022 genes) 174 Rest of genome including XMs (13,804 genes) –All individual siRNA oligos available for retests of hits 4 individual oligos per gene High-Throughput siRNA assays performed in 96- or 384-well plates (384-well preferred) Mouse siRNA library from Qiagen formatted in 384-well plates. Agilent system (96/384 well) –6 stackers –Bravo 96 or 384 tip pipetting head- 9 plate positions –Thermal Plate Sealer –Wellmate Reagent Dispenser (96/384 well) –ELx405 Plate Washer (96/384 well)

High-Content Cellular Imaging ImageXpress Micro (Molecular Devices) –Fully automated inverted epifluorescence microscope scans 6- to 384-well plates and records multiple fluorescent images –Live cell option-Temp, CO 2, humidity –Xenon Lamp/filter based –Complete spectral range with 15 filter set combinations –Up to four color imaging/any fluorophore –Phase Contrast and Brightfield Robotic Arm Integration –3 stackers for fixed plates –Automated incubator for imaging up to 42 live- cell plates Image Analysis Software –MetaXpress 5 with 10 Analysis modules –2 analysis computers –3 rd party/custom/ Matlab, Cell Profiler Completed fully automated whole genome siRNA High Content Screen –200 X 384-well plates imaged, 1 site, 4 colors in 10 weeks; 307,000 images! More Info: David E. Solow-Cordero, Ph.D. Director, HTBC, CCSR-0133