• 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--
R eporter 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)
– 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)

• 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)

• More Info: http://htbc.stanford.edu
David E. Solow-Cordero, Ph.D.
Director, HTBC, CCSR-0133 desolow@stanford.edu
– 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
– 3 stackers for fixed plates
–
Automated incubator for imaging up to 42 livecell plates
– MetaXpress 5 with 10 Analysis modules
– 2 analysis computers
– 3 rd party/custom/ Matlab, Cell Profiler
– 200 X 384-well plates imaged, 1 site, 4 colors in
10 weeks; 307,000 images!