1. Packard BioChip Technologies PerkinElmer Life Sciences 1 Dr. Steve Hawkins Technology Manager The GeneTAC TM G 3

2. Packard BioChip Technologies PerkinElmer Life Sciences 2 Micro-arraying Library management Change the tool to change the function Gridding / replicating Picking Takes less than 10 minutes

3. Packard BioChip Technologies PerkinElmer Life Sciences 3 A common user interface A standard method definition style Simplicity

4. Packard BioChip Technologies PerkinElmer Life Sciences 4 GeneTAC G3 Library Management System A multifunctional tool combining effective genomic library generation and management with array production GeneTAC micro-arrayer G3 –up to 4800spots (10 x 10 x 48) (equivalent to 76800 oligo spots) –Spot size ~300 µm –32 or 48 solid pins –Positional accuracy 100 µm –Positional precision 10 µm –384 well plates –multi-functional capabilities

5. Packard BioChip Technologies PerkinElmer Life Sciences 5 The G3 uses a dip and print technique for solid pins GeneTAC G3

6. Packard BioChip Technologies PerkinElmer Life Sciences 6 Fast cycle time Optimised motion for reproducible transfer of liquids

7. Packard BioChip Technologies PerkinElmer Life Sciences 7 32 or 48 pin tool using solid Titanium pins Printing BioChips

8. Packard BioChip Technologies PerkinElmer Life Sciences 8 Slide holder Marker plate holdercleaning Marker genes are separate from the library

9. Packard BioChip Technologies PerkinElmer Life Sciences 9 Genes spotted: 1920 Probes: (1) Cy3 labeled cDNA probe prepared from Human Adult total RNA (2) Cy5 labeled cDNA probe generated from Human Fetal total RNA Marker:Q gene from Lambda Example

10. Packard BioChip Technologies PerkinElmer Life Sciences 10 Effective Probe DNA Delivery

11. Packard BioChip Technologies PerkinElmer Life Sciences 11 Tracability

12. Packard BioChip Technologies PerkinElmer Life Sciences 12 GeneMAP™ Mouse Clone Array GeneMAP™ Array Chips

13. Packard BioChip Technologies PerkinElmer Life Sciences 13 rapid and accurate Colony picking

14. Packard BioChip Technologies PerkinElmer Life Sciences 14 Can use 96 well, 384 well or agar plates Colony Picking

15. Packard BioChip Technologies PerkinElmer Life Sciences 15 Programmable number of ‘dips’ Clones arranged in known order and position Colony Picking

16. Packard BioChip Technologies PerkinElmer Life Sciences 16 Compatible plate formats Input –15cm Petri –10cm Petri –22cm Bio-Assay –10cm Bio-Assay –GS Culture dish Output –96 well plate –96 deep well –384 well plate –GS Culture dish

17. Packard BioChip Technologies PerkinElmer Life Sciences 17 Plate selection

18. Packard BioChip Technologies PerkinElmer Life Sciences 18 Plate selection Using GS Culture plates Using Bioassay input plates Using Round Petri dishes

19. Packard BioChip Technologies PerkinElmer Life Sciences 19 Brushing to remove agar Sonication to sterilise Sterilization

20. Packard BioChip Technologies PerkinElmer Life Sciences 20 Ethanol for final sterilisation Heat lamp to dry Sterilization

21. Packard BioChip Technologies PerkinElmer Life Sciences 21 define the time for each step using arrows Sterilization Protocol

22. Packard BioChip Technologies PerkinElmer Life Sciences 22 Imaging On-board CCD Camera –Optimised imaging –440,000 pixels on the chip –each pixel represents about 0.01 mm 2 –8 bit grey scale resolution –excellent blue / white discrimination

23. Packard BioChip Technologies PerkinElmer Life Sciences 23 The IP Settings window Example of the Grabbed Image Image processing - IP Edge detect Size and shape Circularity Colour

24. Packard BioChip Technologies PerkinElmer Life Sciences 24 Workflow Picking performance –Typical capacity up to 20,000 clones per 8 hour day up to 15,000 clones per 8 hour day with double inoculation –colonies between 0.5 and 4.0mm –accuracy better than 99% –select on basis of colour, size and shape

25. Packard BioChip Technologies PerkinElmer Life Sciences 25 Gridding onto Hybond-N+ nitrocellulose filter

26. Packard BioChip Technologies PerkinElmer Life Sciences 26 Keeping track custom patterns allow easy cross referencing of the hybridisation results to the corresponding 384-well plate without the use of ink gridding

27. Packard BioChip Technologies PerkinElmer Life Sciences 27 Replication Direct copy of either 96 or 384 well plates using the gridding tool Expand 1 x 384 well to 4 x 96 Contract 4 x 96 to 1 x384 Eliminate the effects of freeze-thaw cycles Fast and accurate copying of libraries

28. Packard BioChip Technologies PerkinElmer Life Sciences 28 Library management Selectively re-array individual elements from a library Addresses individual wells in any library plate Rapid set-up via standard spreadsheet Full report generated Optional autoloader for total plate management Replication