KingFisher Purification: FFPE and cfDNA
undefinedKingFisher Purification: FFPE and cfDNA
June 19
th
, 2018
Kingfisher Duo – Principle and consumables
Principle – Move the beads, not the solutions
96-well plate
24-well plate
Tip combs + magnet
Custom reagent into wells
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Hardware
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The instrument itself
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Software
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Runs BindIt scripts (.bdz files)
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BindIt SW allows users to create new scripts
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Consumables
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Plates
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Tip Combs
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Very Simple Operation
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Turn On Instrument
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Select Protocol
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Load Plate(s)
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Hit “Start”
Uploading and running scripts on the instrument (2 ways):
1.
Use a thumb drive to transfer files directly to the instrument
2.
Transfer from the laptop connected to the instrument (if applicable)
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Automated Purification
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Improved workflow
•
Reduced hands-on time
•
More walkaway time = time better spent on other activities
•
Reduced variability
•
Remove operator-to-operator variability
•
Reproducible, consistent results
•
Compact footprint will fit into any lab
•
40 x 46 x 35 cm
•
Will fit into almost any lab setting, easy to move, install
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Easy to use
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Select program, load plates, hit start
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No advanced training required
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Low price point
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Scalable
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Can run 1-12 samples
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Increase to higher throughput on Flex
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Barcode reader for sample tracking (optional)
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Sample traceability for clinical labs
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Higher throughput processing
capability - One plate at a time
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We do not have agreement in the
works to sell this… yet
•
Scripts written for the Duo will
seamlessly transfer to the Flex for
higher throughput
•
If a customer has a Flex – we can
target them
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Automated DNA and RNA purification from FFPE Samples
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Protocols available on the Covaris website:
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https://covaris.com/wp-content/uploads/pn_010445.pdf
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https://covaris.com/wp-content/uploads/pn_010451.pdf
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Detailed instructions on KingFisher-based DNA and RNA purification
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KingFisher Scripts:
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Each script is a BindIt file
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Separate scripts for FFPE tNA and cfDNA purification
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Available on the website
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Automated cfDNA purification from 1-4mL of plasma
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Available on the website:
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http://covaris.com/truxtrac-cfdna/
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http://covaris.com/wp-content/uploads/pn_010443.pdf
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Detailed instructions on how to run the 1-4mL on the KingFisher Duo
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Scripts:
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Each script is a BindIt file
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Separate scripts for 1mL, 2mL, 3mL, and 4mL processing
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Available as a zip file on the website
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cfDNA
FFPE
Workflow improvements and equivalence data snapshot
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cfDNA and tNA demonstrated equivalence to manual purification
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Workflow improvement
FFPE tNA
cfDNA
cfDNA KF Data
Normalized yield measured by HS dsDNA Qubit Assay
Yields by qPCR
DNA Integrity Index
FA Traces
cfDNA Processing on KF Duo
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Automated cfDNA purification on the Kingfisher Duo
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Dramatic workflow improvement
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Demonstrated equivalence to
manual extraction
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Process 1-4 mLs of plasma
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Process 6 samples at a time in a
24-well plate on a Duo
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Kingfisher Flex: Ability to process
24 samples at a time
KingFisher Purification system for FFPE and cfDNA offers automated purification, reducing hands-on time and variability. Its operational simplicity and key value propositions make it a versatile solution for labs.
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Presentation Transcript
KingFisher Purification: FFPE and cfDNA June 19th, 2018
Kingfisher Duo Principle and consumables Principle Move the beads, not the solutions 96-well plate Tip combs + magnet 24-well plate Custom reagent into wells Proprietary 2
Components of the KingFisher Duo 1. Hardware The instrument itself 2. Software Runs BindIt scripts (.bdz files) BindIt SW allows users to create new scripts 3. Consumables Plates Tip Combs 3
Operational Simplicity Very Simple Operation Turn On Instrument Select Protocol Load Plate(s) Hit Start Uploading and running scripts on the instrument (2 ways): 1. Use a thumb drive to transfer files directly to the instrument 2. Transfer from the laptop connected to the instrument (if applicable) NOTE All KF Scripts are available online 4
KF Duo Plates, Specs, Applications Plate # Wells Max Volume/Well KingFisher Duo # Samples Processed/Run KingFisher Flex # Samples Processed/Run Applications 24-well 24 5mL 96-well 96 1mL 6 12 24 96 cfDNA (1-4mL) FFPE tNA (m-130/m-500) 5
Key Value Props of KingFisher + AFA Automated Purification Improved workflow Reduced hands-on time More walkaway time = time better spent on other activities Reduced variability Remove operator-to-operator variability Reproducible, consistent results Compact footprint will fit into any lab 40 x 46 x 35 cm Will fit into almost any lab setting, easy to move, install 6
Key Value Props Continued Easy to use Select program, load plates, hit start No advanced training required Low price point Scalable Can run 1-12 samples Increase to higher throughput on Flex Barcode reader for sample tracking (optional) Sample traceability for clinical labs 7
KingFisher Flex Higher throughput processing capability - One plate at a time We do not have agreement in the works to sell this yet Scripts written for the Duo will seamlessly transfer to the Flex for higher throughput If a customer has a Flex we can target them 8
FFPE Addendum and Scripts Automated DNA and RNA purification from FFPE Samples Protocols available on the Covaris website: https://covaris.com/wp-content/uploads/pn_010445.pdf https://covaris.com/wp-content/uploads/pn_010451.pdf Detailed instructions on KingFisher-based DNA and RNA purification KingFisher Scripts: Each script is a BindIt file Separate scripts for FFPE tNA and cfDNA purification Available on the website 9
cfDNA Addendum and Scripts Automated cfDNA purification from 1-4mL of plasma Available on the website: http://covaris.com/truxtrac-cfdna/ http://covaris.com/wp-content/uploads/pn_010443.pdf Detailed instructions on how to run the 1-4mL on the KingFisher Duo Scripts: Each script is a BindIt file Separate scripts for 1mL, 2mL, 3mL, and 4mL processing Available as a zip file on the website 10
KF Processing Times (Purification) cfDNA cfDNA Processing Time 1mL Processing Time 2mL Processing Time 3mL Processing Time 4mL Processing Time Processing Time (min) 37 49 61 74 FFPE FFPE Processing Time RNA DNA Processing Time (min) 73 58 11
Workflow improvements and equivalence data snapshot cfDNA and tNA demonstrated equivalence to manual purification Workflow improvement ug DNA 4.50 4.00 3.50 Hand-on Time to Process 24 Samples (minutes) 3.00 2.50 FFPE tNA 2.00 1.50 Competitor Q (Manual) 1.00 0.50 0.00 DNA Column Bead - King Fisher - 7.5 min Bead - King Fisher - 10 min truXTRAC FFPE (Manual) Kingfisher cfDNA 0 50 100 150 200 250 Paraffin removal/Processing RNA Purification DNA Purification Proprietary 12
cfDNA KF Data Qubit Yields Normalized to ng DNA Eluted/ ml Plasma 9 8 7 ng DNA/ ml Plasma 6 5 4 3 2 1 0 1 ml 2 ml 3 ml 4 ml 1 ml KingFisher Manual KP52846 Normalized yield measured by HS dsDNA Qubit Assay Proprietary 13
Yields by qPCR Normalized cfDNA Extraction Yields Measured by ALU115 Primer qPCR Normalized cfDNA Extraction Yields Measured by ALU247 Primer qPCR 9 4.5 8 4 7 3.5 ng DNA/ ml Plasma ng DNA/ ml Plasma 6 3 5 2.5 4 2 3 1.5 2 1 1 0.5 0 KingFisher 1ml KingFisher 2 ml KingFisher 3ml KingFisher 4 ml Manual 1 ml 0 ALU 115 KingFisher 1ml KingFisher 2 ml KingFisher 3ml KingFisher 4 ml Manual 1 ml KP52846 ALU 247 Proprietary 14
DNA Integrity Index DNA Integrity ALU247/ALU115 0.6 ng eluted ALU247/ ng eluted ALU115 0.5 0.4 0.3 0.2 0.1 0 KingFisher 1ml KingFisher 2 ml KingFisher 3ml KingFisher 4 ml Manual 1 ml KP52846 Proprietary 15
FA Traces Proprietary 16
Automated cfDNA purification on the Kingfisher Duo Dramatic workflow improvement Demonstrated equivalence to manual extraction Process 1-4 mLs of plasma Process 6 samples at a time in a 24-well plate on a Duo Kingfisher Flex: Ability to process 24 samples at a time Proprietary 18
