Transcript X-tractor Gene Introduction
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simple affordable complete
Brant Bassam PhD www.corbettlifescience.com
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Easy to use system Lowest possible cost per extraction Hands-free, walk-away operation High speed + high throughput (96 well) Consolidated chemistry
minimum reagents and kit types work for all sample types
Complete integrated system solution
automation, reagents, consumables, protocols, options
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Bench Top Unit
Work surface enclosed by acrylic hood Dual position Manifold 1. Vacuum to Waste 2. Vacuum to Elute Tip discard chute 4 SBS plate positions 3–8 Reagent Tubs (variable volumes) 25 kg 8-Channel pipetting head
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Under Bench Vacuum/Waste Station
Vacuum chamber coupling Pump coupling High power pump Carbon exhaust filter PolyCap pump filter Vacuum sensor Software regulated -5 to -70 kPa, 80 L/min 19 kg www.corbettlifescience.com
To X-tractor Gene manifold Bottle full sensor Waste bottle 4 L sanitary quick disconnect bottle
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Instrument Options
UV Light
• Irradiates surfaces between runs • Robot arm moves so light reaches all areas • Software controlled • Timed post-run operation • Must be factory fitted
HEPA filter
• Positive pressure clean air supply • Software controlled • Integrated design • Easy filter change • Can be added after install www.corbettlifescience.com
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Easy-to-use Wizard For standard protocols simply:
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. Click-select the columns to extract (
in columns of 8
)
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. Press
Jump to End
button and start run!
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Software
Intuitive color-coded interface mirrors actual deck layout Active screen prompts (where appropriate) For each reagent tub position, volumes needed are calculated for the number of samples to be extracted www.corbettlifescience.com
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Barcode Reader Compatible
Sample names and information can also be imported or exported www.corbettlifescience.com
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Pre- and Post-Run Reports
HTML report (universal format) Provides details of a run Documentation & Traceability (e.g. sample, batch and lot identifiers) Can be directly: Printed Saved E-mailed Exported to Word www.corbettlifescience.com
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Bind-Elute Protocol
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CorProtocols
™
for different applications
Available for web download Each comprises: PDF Protocol (Instructions for Use) X-tractor Gene Run File
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CorProtocol
™
Reagents
DNA Extraction:
P/N 2220 DX Reagent Pack P/N 2228 DXT (Tissue Digest Reagent)
Coming Soon:
Viral Kit
(VX Reagent Pack) www.corbettlifescience.com
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DX Reagents
Abbreviations aid recognition (DXL, DXB, etc)
DX Reagent Pack ( P/N 2220) comprises :
1. DXL 2.
– 3.
– 5.
4.
DXB DXW 6.
7.
DXF E Liquid Digest Reagent DX Digest Enzyme DX Binding Additive Binding Reagent Wash Reagent Final Wash Reagent Elution Reagent P/N 2221 P/N 2222 P/N 2224 P/N 2223 P/N 2225 P/N 2226 P/N 2227
DXT Tissue Digest Reagent
(Reagent is for separate purchase if required)
P/N 2228
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X-tractor Consumable Pack
(standard)
P/N 2270 Sufficient for 480 Extractions
960 x OnCor™ C 200 µL Tips 5 x Square-well Lysis Blocks 60 x Elution Plate 8-Strip Caps 14 5 x Capture Plates 5 x Elution Plates (96 plain tube cluster rack) 5 x Sealing Films
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Lysis Plate
15 Square well design for maximum capacity (2.2 mL/well)
P/N 2271 96 Square-well Lysis Plate
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Capture Plate
16 Standard Plate ~10 µg capacity
P/N 0913 Capture Plate, Low Skirt, 800 µL
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Elution Plate
Custom designed rack for X-tractor Gene Convenient format Grid-referenced lid design Tab-lock on lid Cluster rack design (separate tubes) www.corbettlifescience.com
P/N 2147 Elution Plates, 96 Plain Tube Cluster Rack
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Seals
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P/N 1636 Elution Plate 8-Strip Caps
Silicon press-on type Grid referenced (A –H)
P/N 2280 Sealing Film
Seals and protects any unused portion of a capture plate
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OnCor
™
C Tips
rack design SBS rack with replaceable lid for easy removal and storage between runs Total rack enclosure for security and contamination resistance 19 Fully pocketed rack design Separate pockets for each tip prevents cross contamination, allowing tip re-use during a run • Tip re-use reduces costs • Uses only 1.5 tip racks per 96 extractions
P/N 2299 OnCor™ C 200 µL Fine Bore Tip 96 tips/rack, 10 racks/box
OnCor
™
C Tips
design evolution www.corbettlifescience.com
20 • Unmodified tip Filter set too low • Easily wet and blocked • High-Set Filter Design Better liquid clearance height • Less wetting issues OnCor ™ C Design • Best liquid clearance height • Optimized barrel and taper • Improved filter matrix
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OnCor
™
C Tips
Advanced robotic tip design exclusive to Corbett Robotics
Capp™ compatible hub dimensions (“C” tips) Larger filter and new more effective matrix Optimized terminal taper minimizes sample immersion and displacement effects Fine bore tip for precision liquid handling www.corbettlifescience.com
High-Set filter guards against filter wetting Wide upper barrel for greater liquid capacity and clearance between the liquid and filter inside the tip Low retention polymer for improved liquid shedding and pipetting precision
P/N 2299 OnCor™ C 200 µL Fine Bore Tip 96 tips/rack, 10 racks/box
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OnCor
™
C Tips
improved liquid clearance height www.corbettlifescience.com
Unmodified tip Example wet filter High-Set Filter OnCor ™ C clearance height with 200 µL liquid
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Disposable Reagent Tubs
Convenient Streamlines setup Graduated for volume (mL) Reusable lids curb evaporation Extremely low reagent dead volume (e.g. 0.75 mL in a 70 mL tub) www.corbettlifescience.com
P/N 2137 70 mL size P/N 2136 170 mL size P/N 2314 270 mL size
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CorProtocol
™
Options
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Options
25 High Yield Capture Plate ~40 µg capacity
P/N 1863 High Yield Capture Plate
Tube Options
Volumes: 0.65 mL Standard tubes 0.5 mL Twist-Lock tubes www.corbettlifescience.com
Cap options:
Grid referenced tubes in racks with individual Twist-Lock caps* Silicon 8-strip caps
P/N 1636 26
Label options:
1.
Grid Reference* 2.
3.
2D Cipher Barcode* Plain Unlabelled
( P/N 2147 in purple rack, not shown ) H9 * from ABgene or authorized distributor only
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rd
Party Chemistry Options
Accessories are available to use a wide range of kits www.corbettlifescience.com
27 Please note Corbett Robotics cannot provide support for reagents and plasticware of other manufacturers
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Sample Digest Option
Vortemp 56 Shaking Incubator Can improve speed & completeness of digest Variable vortex speed —200 to 1500 rpm Broad temperature range to 100 °C Interchangeable platforms for Microtubes and Microplates www.corbettlifescience.com
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DX Reagent Verification Testing Experimental setup
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Reagents and Consumables
DX Reagent Pack (P/N2220) was functionally tested using Whole Blood CorProtocol™ standard X-tractor Consumable Pack (standard; P/N 2270).
A B C D E F G H
Samples
Whole bovine blood was collected in an EDTA Vaccutainer Maximum whole blood sample size was tested (200 µL) 96 samples total were processed (whole 96-well plate); 48 whole blood and 48 no template controls (NTCs) To test for well-to-well cross contamination, samples were extracted in a checkerboard pattern (as shown) alternating positive samples ( ● ) and NTCs ( ● ).
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DX Reagent Verification Testing
cont.
Gel electrophoresis of raw extracted DNA from positive samples:
reproducible yields of high quality, intact high molecular weight DNA (~40 kb) MW B1 D1 F1 H1 A2 C2 E2 G2 B3 D3 F3 H3 A4 C4 E4 G4 B5 D5 F5 H5 A6 C6 E6 G6 MW 40 Kb 20 Kb 10 Kb 6 Kb 4 Kb 3 Kb 2 Kb 1 Kb MW B7 D7 F7 H7 A8 C8 E8 G8 B9 D9 F9 H9 A10 C10 E10 G10 B11 D11 F11 H11 A12 C12 E12 G12 MW 40 Kb 20 Kb 10 Kb 6 Kb 4 Kb 3 Kb 2 Kb 1 Kb
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DX Reagent Verification Testing
cont.
Spectrophotometer analysis of all positive samples
2.000
1.800
1.600
1.400
1.200
1.000
0.800
0.600
0.400
0.200
0.000
0 8
260/280 Ratio
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Sample #
32 40 48 90 80 70 60 50 40 30 20 10 0 0
DNA Concentration ng/µL (from 180 µL whole blood eluted in 150 µL)
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Sample #
32 40 48
Sample Maximum Minimum dsDNA Yield
9.39 µg 7.26 µg 8.41 µg
Vol. Yield
125 µL 119 µL 121.7 µL
Concentration
77.0 ng/ µL 60.0 ng/ µL 69.1 ng/ µL
A260/280 Ratio
1.89
1.84
Average
1.87
Std. Dev.
0.52 µg 1.59 µL 4.4 ng/ µL 0.01
Note: the results shown are indicative only, actual yields will vary depending on the species and the health of the individual from which blood was harvested.
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DX Reagent Verification Testing
cont.
Real-time amplification analysis of all samples
0.25
0.20
DX Reagent Verification Run ■ Positive samples ■ NTC samples 0.15
0.10
Maximum C T Minimum C T Average C T C T Std. Dev.
25.04
22.19
24.59
0.18
0.05
Threshold 0.00
5 10 15 20 Cycle 25 30 35 40
Real-Time Analysis
Real-Time quantitative amplification analysis was done on all samples targeting the G6PD1 gene to single copy sensitivity (40 cycles) to test for inhibitors and for evidence of cross-contamination
Amplification Results
• early, reproducible amp. of all positive samples • no evidence of PCR inhibitors
Cross-contamination Results
• no amplification of alternate NTC samples • no evidence of cross-contamination
Using the X-tractor Gene ™ will minimise any lab’s contamination risk!
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Some sample types successfully extracted
Blood whole, fresh, frozen, buffy coat Human, Mouse, Turtle, Bovine… Serum/Plasma, HIV, Hep B , Hep C, CMV Urine Chlamydia and N. gonorrhoea Swabs HSV1 and 2 Tissue Mouse Tails Prawn Tissue Bird Quill Pulp Nasal Aspirates MPV (a single stranded RNA Virus) Cultured cells Micro Organisms Bacteria (Gram Pos. and Gram Neg.) and Fungi
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Benefits
Consistency & Workflow Reduce human error and free-up technician time Standardisation Between labs and runs Documentation Pre- and Post-Run reports Safety No phenol/chloroform All waste safely contained for disposal Harmful fumes are carbon filtered out Work area completely enclosed by hood Run pauses if the hood is opened www.corbettlifescience.com
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Summary Complete-Solution
Instrument, reagents, consumables, protocols
Substantial Cost Reduction
About half other methods
Minimum Possible Cross-Contamination Risk
Optimized liquid handling strategies
High-Throughput
96 samples in about 1.5 hours
Versatility and Choice
Compatible with a wide range of protocols and 3 rd party chemistries
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96-well Workflow Integration
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Gene Disc™ 100
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Offices
Brisbane Australia
Corbett Robotics Pty Ltd 42 McKechnie Drive Eight Mile Plains, QLD 4113 T +61 7 3841 7077 F +61 7 3841 6077
Sydney Australia
Corbett Research Pty Ltd 14 Hilly Street Mortlake, NSW 2137 T +61 2 9736 1320 F +61 2 9736 1364
United Kingdom
Corbett Research UK Ltd Unit 1 Terek House, Sandpiper Court Phoenix Business Park, Eaton Socon St Neots, Cambridgeshire, PE19 8EP T +44 (0) 1480 407222 F +44 (0) 1480 407999
USA
Corbett Robotics Inc 185 Berry Street, Suite 5200 San Francisco, CA 94107 USA T +1 415 348 1166 F +1 415 348 1177
Web
www.corbettlifescience.com
All slides 2006 Corbett Life Science. All rights reserved