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

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PackardBioChip TechnologiesPerkinElmer Life Sciences

Dr. Steve HawkinsTechnology Manager

The GeneTACTM G3

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Micro-arraying

Library management

Change the tool to change the function

Gridding / replicating

Picking

Takes less than 10 minutes

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• A common user interface• A standard method definition style

Simplicity

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• GeneTAC G3 Library Management SystemA 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

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The G3 uses a dip and print technique for solid pins

GeneTAC G3

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Fast cycle time

• Optimised motion for reproducible transfer of liquids

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32 or 48 pin tool using solid Titanium pins

Printing BioChips

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Slide holder

Marker plate holdercleaning

Marker genes are separate from the library

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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

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Effective Probe DNA Delivery

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Tracability

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GeneMAP™ Mouse Clone Array GeneMAP™ Array Chips

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rapid and accurate

Colony picking

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Can use 96 well, 384 well or agar plates

Colony Picking

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Programmable number of ‘dips’Clones arranged in known order and position

Colony Picking

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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

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Plate selection

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Plate selection

Using GS Culture plates

Using Bioassay input plates

Using Round Petri dishes

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•Brushing to remove agar•Sonication to sterilise

Sterilization

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•Ethanol for final sterilisation•Heat lamp to dry

Sterilization

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define the time for each step using arrows

Sterilization Protocol

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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

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The IP Settings window Example of the Grabbed Image

Image processing - IP

Edge detectSize and shape

CircularityColour

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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

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Gridding onto Hybond-N+ nitrocellulose filter

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Keeping track

• custom patterns allow easy cross referencing of the hybridisation results to the corresponding 384-well plate without the use of ink gridding

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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

Master 5

Master 6

Master 7

Master 8

Master 3

Master 4

Master 2

Master 1

Second TrayUser promptedwhen to change

Copy 8a

Copy 8bCopy 6b

Copy 6a

Copy 4aCopy 2a

Copy 4bCopy 2b

Copy 3a

Copy 3b

Copy 1a

Copy 1b

Copy 7a

Copy 7bCopy 5b

Copy 5a

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Library management Selectively re-array individual

elements from a library Addresses individual wells in any

library plate

Rapid set-up via standard spreadsheetFull report generatedOptional autoloader for total plate management

Replication

Documents

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