Automated Purification System: KingFisher Flex & Duo

Explore the KingFisher automated purification system featuring the Duo and Flex models. Learn about the principles, components, operational simplicity, plate specs, applications, and key value propositions. Discover how these systems offer improved workflow, reduced hands-on time, scalability, and reproducible results in a compact design. The KingFisher Flex allows for higher throughput processing with seamless script transfer from the Duo.

• Automated purification
• KingFisher Flex
• KingFisher Duo
• Workflow efficiency
• Reproducible results

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1. KingFisher Purification: FFPE and cfDNA June 19th, 2018

2. 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

3. 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

4. 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

5. 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

6. 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

7. 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

8. 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

9. 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

10. 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

11. 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

12. 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

13. 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

14. 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

15. 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

16. FA Traces Proprietary 16

17. cfDNA Processing on KF Duo 17

18. 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

19. cfDNA Plate Map Reduce risk of error for reagent loading Proprietary 19

20. Detailed Processing Times on KF Duo Proprietary 20

21. FFPE 12 Samples (Purification times not final) 12 samples (based on processing with an L transducer) DNA AFA 1 PK 60 Decross 60 Purify 75 Total 271 Step Minutes AFA 15 PK 20 Centrifuge 20 RNA Decross 20 Purify 75 Total 150 12 samples (based on processing with an E transducer) DNA AFA 1 PK 60 Decross 60 Purify 75 Total 326 Step Minutes AFA 70 PK 20 Centrifuge 20 RNA Decross 20 Purify 75 Total 205 21

22. FFPE 24 Samples (Purification times not final) 24 Samples (based on processing with an L transducer) DNA AFA 1 PK 60 Decross 60 Purify 1st 12 Purify 2nd 12 75 Total 351 60 Step Minutes AFA 35 PK 20 Centrifuge 20 RNA Decross 20 Purify 1st 12 Purify 2nd 12 75 Total 230 60 24 Samples (based on processing with an E transducer) DNA AFA 1 PK 60 Decross 60 Purify 1st 12 Purify 2nd 12 75 Total 456 60 Step Minutes AFA 140 PK 20 Centrifuge 20 RNA Decross 20 Purify 1st 12 Purify 2nd 12 75 Total 335 60 22

23. cfDNA - 1mL (L series) 1mL (processing with an L transducer) 6 Samples Step Minutes PK 20 AFA 7 Purify 40 Total (6) 67 12 Samples PK 20 AFA 7 Purify 40 Total (12) 94 18 Samples PK 20 AFA 7 Purify 40 Total (18) 121 24 Samples PK 20 AFA 7 Purify 40 Total (24) 148 23

24. cfDNA - 1mL (M or E series) 1mL (processing with an E transducer) 6 Samples Step Minutes PK 20 AFA 15 Purify 40 Total (6) 75 12 Samples PK 20 AFA 15 Purify 40 Total (12) 110 18 Samples PK 20 AFA 15 Purify 40 Total (18) 145 24 Samples PK 20 AFA 15 Purify 40 Total (24) 180 24

25. cfDNA - 4mL (L series) 4mL (based on processing with an L transducer) 6 Samples Step Minutes PK 20 AFA 15 Purify 80 Total (6) 115 12 Samples PK 20 AFA 15 Purify 80 Total (12) 150 18 Samples PK 20 AFA 15 Purify 80 Total (18) 185 24 Samples PK 20 AFA 15 Purify 80 Total (24) 220 25

26. cfDNA - 4mL (M or E series) 4mL (processing with an E transducer) 6 Samples Step Minutes PK 20 AFA 55 Purify 80 Total (6) 155 12 Samples PK 20 AFA 55 Purify 80 Total (12) 230 18 Samples PK 20 AFA 55 Purify 80 Total (18) 305 24 Samples PK 20 AFA 55 Purify 80 Total (24) 380 26