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Sample & Assay Technologies - 1 - smart compact affordable Brant Bassam PhD QIAxtractor Andrea Tesoriero Application Specialist.

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Presentation on theme: "Sample & Assay Technologies - 1 - smart compact affordable Brant Bassam PhD QIAxtractor Andrea Tesoriero Application Specialist."— Presentation transcript:

1 Sample & Assay Technologies smart compact affordable Brant Bassam PhD QIAxtractor Andrea Tesoriero Application Specialist

2 Sample & Assay Technologies QIAxtractor is a seamless integration to your workflow post PCR analysis real time PCR or RT PCR PCR setup nucleic acid extraction Sample preparation PCR & analysis PCR setup enzymatic digestion post purification QC enzymatic digestion aliquoting DNA extraction Sample preparation

3 Sample & Assay Technologies COMPACT QIAxtractor moves according to the experiment requirements Fits into any work space and is easily movable Very small footprint Light and easily movable (25 kg; 51 lbs) Vacuum and waste stations located under the bench for more space and convenience 8-channel pipetting head without any tubing for more accuracy and easy maintenance Comes with a laptop for even more flexibility 8-Channel pipetting head

4 Sample & Assay Technologies Bind-Elute Protocol Sample loaded with lysis buffer at 1:2 ratio Lyse for 10 minutes Load the capture plate and apply vacuum Wash captured DNA with 1 Lysis Buffer 2 Propanol wash 1 Ethanolic wash Residual Ethanol is removed by vacuum Capture plate moves to the elution station Elution buffer loaded and vacuum applied DNA elutes into a 96-well cluster tube plate

5 Sample & Assay Technologies ̣ Minimize cross contamination risk ̣ Reduce human error ̣ Reduce costs ̣ Increase throughput & save time Often the bottleneck in a project Work in increments of 8 samples (from 8 to 96) 96 samples in about 96 minutes ̣ Increase safety Minimum exposure to dangerous chemicals ̣ Versatility and choice DX liquid or tissue reagent, Vx reagent Wide range of protocols for DNA, RNA and viral nucleic acids Associated plasticware ̣ Maintain your sanity (pain threshold is about 25 extractions per week!) Why automate?

6 Sample & Assay Technologies Sample types successfully extracted ̣ Blood -whole, fresh, frozen, buffycoat,Human, Mouse, Turtle, chickens, sheep Bovine Serum/Plasma, -HIV, Hep B, Hep C, CMV ̣ Urine Chlamydia and N. Gonorrhoea ̣ Swabs HSV1 and 2 ̣ Solid 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 ̣ Plants Fruit, bark, leaves and stems ̣ Faeces Norris Virus ̣ Forensics cigarette butts, food scraps, finger prints, hair, fabric, FTA cards

7 Sample & Assay Technologies ̣ Easy to use system ̣ Low cost 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 QIAxtractor

8 Sample & Assay Technologies mm 390 mm closed 770 mm open 650 mm 25 kg Bench Top Unit 4 SBS plate positions Tip discard chute 3–8 Reagent Tubs (variable volumes) Work surface enclosed by transparent cover Dual position Manifold 1. Vacuum to Waste 2. Vacuum to Elute Channel pipetting head

9 Sample & Assay Technologies kg 520 mm 495 mm 400 mm Under Bench Vacuum/Waste Station Waste bottle 4 L sanitary quick disconnect bottle Bottle full sensor Vacuum sensor Software regulated -5 to -70 kPa, 80 L/min PolyCap pump filter Carbon exhaust filter Pump coupling Vacuum chamber coupling High power pump To X-tractor Gene manifold

10 Sample & Assay Technologies UV Light Irradiates surfaces between runs Robot arm moves so light reaches all areas Software controlled Timed post-run operation Must be factory fitted Instrument Options HEPA filter Positive pressure clean air supply Software controlled Integrated design Easy filter change Can be added after install

11 Sample & Assay Technologies CorProtocols for different applications Available for web download Each comprises: PDF Protocol (Instructions for Use) X-tractor Gene Run File

12 Sample & Assay Technologies Easy-to-use Wizard For standard protocols simply: 2.Press Jump to End button and start run! 1.Click-select the columns to extract (in columns of 8)

13 Sample & Assay Technologies Software For each reagent tub position, volumes needed are calculated for the number of samples to be extracted Intuitive color-coded interface mirrors actual deck layout Active screen prompts

14 Sample & Assay Technologies Barcode Reader Compatible Sample names and information can also be imported or exported

15 Sample & Assay Technologies 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 ed Exported to Word

16 Sample & Assay Technologies X-tractor Plasticware Pack (standard) 5 x Capture Plates 5 x Square-well Lysis Blocks P/N 2270 Sufficient for 480 Extractions 960 x OnCor C 200 µL Tips 5 x Elution Plates (96 plain tube cluster rack) 60 x Elution Plate 8-Strip Caps 5 x Sealing Films

17 Sample & Assay Technologies 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 P/N 2299 OnCor C 200 µL Fine Bore Tip 96 tips/rack, 10 racks/box 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

18 Sample & Assay Technologies QIAxtractor minimizes cross-contamination risk Real-time amplification analysis of all samples 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 samples no evidence of PCR inhibitors Cross-contamination Results no amplification of NTC samples no evidence of cross-contamination

19 Sample & Assay Technologies QIAxtractor optimizes nucleic acid extraction yield Application in Veterinary Diagnostics Results significant higher concentrations of DNA recovered after automated extraction (p=0.001) significant stronger Ct values for the automated solution (p=0.01) Real time analysis evaluation of DNA quality: detection of the universal 18S rRNA housekeeping gene detection of EHV-1: amplification of the glycoprotein B (gB) gene QIAxtractor (DX) Manual method (DNeasy) DNA concentration from blood aliquots (ug/100ul) [Absorbance at 260nm] 5.56 (+/- 3.98)2.44 (+/- 1.21) Cycle threshold (Ct) values [threshold=0.1, baseline 3-15] 21.1 (0.4)22.2 (0.5) Comparison of extraction efficiencies and DNA quality between manual and automated DNA extraction methods by real time analysis Viral detection of EHV-1 horses Dr. N. Pusterla at University of California, School of Veterinary Medicine

20 Sample & Assay Technologies Rapid & sensitive detection of large numbers of specimens Application in Medical Diagnostics Manual: QIAamp viral RNA Manual: Roche high pure viral RNA Universal + QIAamp Virus Biorobot MDX QIAxtractor + VX chemistry Time for 36 specimens 2h -- Time for 96 specimens --2h301h30 Time Operator was actually busy 2h 25 min15 min Experiment: total RNA extraction from fecal specimens + Analysis by multiplex RT PCR Detection of noroviruses in fecal specimens Dr. J.A. Marshall at Victorian Infectious Diseases Reference Laboratory, Victoria (Australia) Results no significant difference between the manual and automation methods automation methods required less material automation methods process more samples in less time

21 Sample & Assay Technologies 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

22 Sample & Assay Technologies SMART QIAxtractor is as convenient as QIAGEN can be Fast 96 samples in 96 min Membrane tolerance: double flow rate; 4x more binding capacity compared to the other plates on the market Robust Just 2 chemistries (DX/VX) for NA extraction from virtually any sample type Consolidated chemistry for liquids and tissues Ultra-effective lyses buffer Easy Very intuitive and flexible software Just a few clicks of the mouse are enough to start a run Safe Innovative features: enclosed hood, UV light, HEPA filter … Optimal protection of the extraction and of the operator Individual elution tubes for easy downstream processing Re-use of tips results in little waste

23 Sample & Assay Technologies ̣ Gracias por su atención

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