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Dr. Steve Hawkins Technology Manager

The GeneTAC TM G 3. Dr. Steve Hawkins Technology Manager. Takes less than 10 minutes. Gridding / replicating. Picking. Library management. Micro-arraying. Change the tool to change the function. Simplicity. A common user interface A standard method definition style.

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Dr. Steve Hawkins Technology Manager

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  1. The GeneTACTM G3 Dr. Steve Hawkins Technology Manager

  2. Takes less than 10 minutes Gridding / replicating Picking Library management Micro-arraying Change the tool to change the function

  3. Simplicity • A common user interface • A standard method definition style

  4. GeneTAC micro-arrayer G3 • GeneTAC G3 Library Management System A multifunctional tool combining effective genomic library generation and management with array production • 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. The G3 uses a dip and print technique for solid pins GeneTAC G3

  6. Fast cycle time • Optimised motion for reproducible transfer of liquids

  7. Printing BioChips 32 or 48 pin tool using solid Titanium pins

  8. cleaning Marker plate holder Slide holder Marker genes are separate from the library

  9. Example 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

  10. Effective Probe DNA Delivery

  11. Tracability

  12. GeneMAP™ Array Chips GeneMAP™ Mouse Clone Array

  13. Colony picking rapid and accurate

  14. Colony Picking Can use 96 well, 384 well or agar plates

  15. Colony Picking Programmable number of ‘dips’ Clones arranged in known order and position

  16. 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 Compatible plate formats

  17. Plate selection

  18. Using GS Culture plates Using Bioassay input plates Using Round Petri dishes Plate selection

  19. Sterilization • Brushing to remove agar • Sonication to sterilise

  20. Sterilization • Ethanol for final sterilisation • Heat lamp to dry

  21. define the time for each step using arrows Sterilization Protocol

  22. Imaging • On-board CCD Camera • Optimised imaging • 440,000 pixels on the chip • each pixel represents about 0.01 mm2 • 8 bit grey scale resolution • excellent blue / white discrimination

  23. Size and shape Circularity Colour Example of the Grabbed Image Edge detect Image processing - IP The IP Settings window

  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. Gridding onto Hybond-N+ nitrocellulose filter

  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. 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. Replication 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

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