MagNA Pure 96 integration in an automatic workflow
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1 MagNA Pure 96 integration in an automatic workflow Dr. Franco Bertolucci
2 Preface Current Challenges for Laboratories Driven by a shift from conventional methods to molecular analysis Increasing need to improve lab efficiency Questions: How can I (re-)organize my lab with available resources? -> Automate time-intensive routine tasks How do I keep up with modern quality management (e.g., documentation obligations)? -> Implement applications as workflows Data Workflow Sample Registration Sample Distribution/ Pre-Prep Sample Purification Setup Downstream Analysis Downstream Analysis Experimental Workflow
3 RAS Workflow Integration With MagNA Pure 96 SW 2.0 and post-elution pipetting Hamilton MagNA Starlet MagNA Pure 96 Light Cycler 480 PCR Set-Up Primary sample Purification Real-Time PCR Sample-in File sharing Result-out From sample to result in 2,5 hours (20 min / 60 min / 50 min) No manual re-entry of data
4 MagNA Pure 96 System Revolutionized Automated Nucleic Acid Isolation 96 Standard Samples < 1h Convenience Roche Quality Versatile applications Run Surveillance/Tracking (FDA compliant) Fully automated Connectivity 4
5 MagNA Pure 96 System Proven Magnetic Particle Technology 12 year expertise in development and application Reagents manufactured with Roche quality Eluted nucleic acid has a reproducibly high standard regarding recovery, integrity and purity 5
6 MagNA Pure solutions for automatic nucleic acids purification More than 10 years of evolution Minutes Minutes Minutes Minutes Samples Channels Duration
7 Isolation Technology Tube Separation Sample Cell disruption Binding Separation Removal of debris Elution Lysis buffer of NA (magnetism) by extensive washing high temperature Proteinase K to MGP of MGP-NA and subsequent separation low salt complex 7
8 MagNA Pure 96 System High throughput in a new dimension 96 Standard Samples < 1h Benefits: Get in faster time to result Getmoreresultsin thesametime Be prepared for the next pandemic (about 800 samples in 8 h possible) 100% simultaneous handling of your samples Benefit from 100% parallel handling of your samples No waiting times No different treatment of your samples So no danger of artifacts and mistakes due to different treatment Treat all you samples as urgent samples
9 MagNAPure 96 Front view Service Hood Housing Maintenance Flap Status LED Load Flap Power On Button/ Main Switch 9
10 Instrument Stage (Details) 1 = Reagent Head 5 = Process Head 9 = Purification Rack 2 = Wash Station 6 = Slot for Bottle Rack 10 = Cooling Plate 3 = Init Bold Reagent Head 7a/7b = Reagent Racks 11 = Process Station 4 = Tip Park Position 8 = Tip Rack 12 = Waste Rack 10
11 1 Reagent Head Characteristics: Four Fluid Channels. Aspiration and dispensing of fluids, e.g., alcohol, saline buffer, magnetic glass particles. Connecting tube of pump and Loop Unit Micro Gear Pump Reagent Loop Unit Needle Mount Barcode Reader Needle Init Bold
12 3 Init Bold Reagent Head Characteristics: Check of each needle if bent (during initialization). A. Initialization step 1: needle on top of the Init Bold. B. Initialization step 2: needle on one side of the Init Bold.
13 5 Process Head (with Tips) Characteristics (Tips): Aerosol filter included. Handles volumes in the range of 5 1,000 μl. Ejector Tips Characteristics (LPS): Checks Tip Park Position. Checks empty tip trays in the Waste Rack. Drop Catcher +Fleeze Presence of Waste Cover in the Waste Rack. Characteristics (Drop Catcher + Fleeze): Prevents droplets falling from the tips onto the stage. The fleeze absorbs the droplets.
14 5 Process Head Fixing of a Tip to a Nozzle Procedure: Step 1: Attachment of outer part of the nozzle (green) to the rim of the tip and inner part (brown/black) into the tip. Step 2: Plunger (black) compresses the o-ring and fixes the tip. Result: The tip is tightly bound to the nozzle. Step 1 Step 2 Result Hamilton CO-RE technology
15 6 Slot for Bottle Rack / Bottle Rack Characteristics: Bottle Rack is inserted into the slot for Bottle Rack. Bottle Rack is divided into three parts, for: A. Internal Control Tubes; B. DNase Bottles; C. MGP Bottles (shaking of MGP bottles starts ~2 min. prior to dispensing the beads). Internal Control Tube DNase Bottles MGP Bottles (Shaker!) EACH DNase and MGP bottle is fixed with a clip Motor of MGP shaker is located under the Bottle Rack.
16 MagNA Pure 96 System Convenience never seen before Few disposables Loading of drawers Benefits: Minimized hands-on time Less storage place Less waste 16
17 11 Process Station Characteristics: Location beneath the Purification Rack. A. Separation Plate with 24 magnet pins moving below the Sample Cartridge (one pin serves four wells). B. Incubator Plate moving below the Sample Cartridge (elution of nucleic acids at~60 C). 17
18 11 Process Station (Functionality) Characteristics: Process Station is turned upside down (depending on the purification protocol step) to change heating and separation functionality. Process Station can move to both positions for Sample/ Processing Cartridges within the Purification Rack. 18
19 MagNA Pure 96 System Just a few Mouse Clicks Software: Intuitive: with an easy-to-learn interface Ready-to-use: includes preprogrammed purification protocols Additional downloadable applications: free protocol updates Simple template generation: easy importing eliminates redundant data entry
20 Appendix: MagNA Pure 96 System Software Overview Screen 20
21 MagNA Pure 96 System High throughput in a new dimension Technical Features: Simply upload all run data Or use the quick setup mode 21
22 MagNA Pure 96 System Convenience never seen before Prefilled reagent trays Ready to use No further assembly needed Barcode labeled 22
23 MagNA Pure 96 System Convenience never seen before Standard Run: 2 reagent trays (+ MGP bottle, system fluid) 2x 2 processing cartridges 2 sets of 96 tips 2x 2x 1x 1 output plate 23
24 MagNA Pure 96 System Components Low reagent complexity for a large number of applications MP 96 tna / DNA Isolation Kit Small Volume 3 x 192 purifications MP 96 tna / DNA Isolation Kit Large Volume MP 96 RNA Isolation Kit Large Volume 3 x 96 purifications 3 x 96 purifications 24
25 MagNA Pure 96 System Large spectrum of applications Genomic DNA Bacterial DNA Viral DNA & RNA Total RNA From different sample materials : Whole blood, blood cells, white blood cells serum/plasma Tissue Paxgene blood body fluids, urine, fecal samples, sputum, etc. etc. More protocols in development (2010/11) : Universal Viral NA (available) - DNA LargeVol blood (1000μl) (available) Bakteria (available) - RNA from Blut (available) DNA fresh tissue (available) - DNA FFPE
26 Hardware SW2.0 vs. SW1.0/SW1.1 MagNA Pure 96 Plate Holder (# ): The plate holder is optional and ONLY necessary when custom plate types are used. This adapter is needed for placing the specific custom plate types (e.g., the LightCycler 480 Multiwell Plate 96) in the purification rack. Anytime when using custom plates in position 3 of the purification rack, place these plates into the plate holder. 26
27 MagNA Pure 96 Applications Genomic DNA Integrity Genomic DNA from 200 μl whole blood (6600 leukocytes/μl) Nanodrop Analysis: Yield: 6.46 μg, Std Dev OD 260/280: 1.95
28 MagNA Pure 96 Applications Genomic DNA Linearity Eluate Undiluted CP: 17.7; Std. Dev: 0.26 Eluate Dilution 1:10 CP: 20.9; Std. Dev: 0.53 LightCycler Real-time PCR for the Cyclophilin A Gene
29 MagNA Pure 96 Applications Viral DNA and RNA Reproducibility LightCycler Real-Time PCR for Parvo B19 Virus
30 MagNA Pure 96 Applications Viral DNA and RNA No Evidence for Cross-contamination Viral NA 500 μl (Parvo B19 in horse serum)
31 MagNA Pure 96 Applications Total RNA Integrity of RNA 28S rrna 18S rrna MWM III 1.00x10 6 K-562 cells 1.00x10 6 HeLa cells
32 MagNA Pure 96 System Automate time-intensive routine tasks Your benefits: more or faster results Standardization of routine jobs save valuable ressources Highly flexible in terms of sample variations: from low numbers to hundreds (in case of pandemics) data management FDA compatible Trust the number 1 in diagnostics with top level service and support
33 We Innovate Healthcare
34 Appendix: MagNA Pure 96 System Software Workplace Screen Ordering 34
35 Appendix: MagNA Pure 96 System Software Workplace Screen Batch Results 35
36 Appendix: MagNA Pure 96 System Software Utilities Query Function 36
37 Appendix: MagNA Pure 96 System Software Utilities User Administration 37
38 7a/7b Reagent Racks 1 (left) + 2 (right) Characteristics: A. One position for a Processing Cartridge. B. Three positions for Reagent Trays. Barcodes, for detection of empty positions in the rack during the resource check Hardware coding for correct positioning of Processing Cartrigdes (blue) and Reagent Trays (red) Front OG.link Reagent Racks 38
39 8 Tip Rack Characteristics: Four positions for tip trays. Barcodes, for detection of empty positions in the rack during the resource check Barcodes of the tip trays on the left hand side. Front OG.link Tip Rack 39
40 9 Purification Rack / 10 Cooling Plate Characteristics: Nucleic acid isolation is performed in this rack. A. Position for the Sample Cartridge and the Process Station. B. Position for an additional Processing Cartridge and the Process Station (necessary for specific protocols). C. Position for Custom Plate (= customer specific Output Plate). D. Position for MagNA Pure 96 Output Plate. Barcodes, for detection of empty positions in the rack during the resource check Front Front OG.link Purification Rack 40
41 12 Waste Rack Characteristics: Rack for liquid waste and used tips. A. Position for Waste Cover. B. Three positions for empty tip trays. Waste Cover Barcodes, for detection of empty positions in the rack during the resource check Barcodes of the empty tip trays on the right hand side (this is the ONLY rack where the barcodes are positioned on the right hand side). Front Front Front OG.link Waste Rack 41
42 Liquid Waste Funnel Characteristics: Placed under the rearmost position of the Waste Rack. Transport of liquid waste via a tube into the Waste Container. Located under the Waste Cover. Liquid Waste Funnel Front Front Front Waste Rack OG.link Liquid Waste Funnel 42
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