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QIAxtractor. smart compact affordable . Brant Bassam PhD. Andrea Tesoriero Application Specialist. Sample preparation. nucleic acid extraction. PCR setup. real time PCR or RT PCR. post PCR analysis . QIAxtractor is a seamless integration to your workflow.
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QIAxtractor smart compact affordable Brant Bassam PhD Andrea Tesoriero Application Specialist
Samplepreparation nucleic acidextraction PCR setup real time PCR or RT PCR post PCR analysis QIAxtractor is a seamless integration to your workflow Samplepreparation DNAextraction aliquoting enzymaticdigestion post purification QC enzymaticdigestion PCRsetup PCR & analysis
COMPACTQIAxtractor 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-Channelpipetting head
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 4
Minimize cross contamination risk Reduce human error Reduce costs Increase throughput & save time Often the bottleneck in a projectWork 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? 5
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 6
Easy to use system Low cost possible cost per extraction Hands-free, walk-away operation High speed + high throughput (96 well) Consolidated chemistryminimum reagents and kit types work for all sample types Complete integrated system solutionautomation, reagents, consumables, protocols, options QIAxtractor 7
Bench Top Unit Work surface enclosed by transparent cover 770 mm open Dual position Manifold 1. Vacuum to Waste 2. Vacuum to Elute 2 2 1 1 Tip discard chute 8-Channelpipetting head 390 mm closed 4 SBS plate positions 3–8 Reagent Tubs(variable volumes) 650 mm 490 mm 25 kg 8
Under BenchVacuum/Waste Station To X-tractor Gene manifold Vacuum chamber coupling Bottle full sensor Pump coupling High power pump 520 mm Waste bottle4 L sanitary quick disconnect bottle Carbon exhaust filter PolyCap pump filter Vacuum sensor Software regulated -5 to -70 kPa, 80 L/min 495 mm 400 mm 19 kg 9
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 10
CorProtocols™ for different applications PDF Protocol (Instructions for Use) X-tractor Gene Run File • Available for web download • Each comprises: 11
Easy-to-use Wizard For standard protocols simply: 1. Click-select the columns to extract(in columns of 8) 2. Press Jump to End button and start run! 12
Software • Intuitive color-coded interface mirrors actual deck layout Active screen prompts For each reagent tub position, volumes needed are calculated for the number of samples to be extracted 13
Barcode Reader Compatible • Sample names and information can also be imported or exported 14
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 15
X-tractor Plasticware Pack(standard) 60 x Elution Plate 8-Strip Caps 960 x OnCor™ C 200 µL Tips 5 x Square-well Lysis Blocks 5 x Capture Plates 5 x Elution Plates(96 plain tube cluster rack) 5 x Sealing Films P/N 2270 Sufficient for 480 Extractions 16
OnCor™ C Tips rack design • 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 • 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 17
QIAxtractor minimizes cross-contamination riskReal-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
QIAxtractor optimizes nucleic acid extraction yieldApplication in Veterinary Diagnostics Viral detection of EHV-1 horsesDr. N. Pusterla at University of California,School of Veterinary Medicine Comparison of extraction efficiencies and DNA qualitybetween manual and automated DNA extraction methods by real time analysis • 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 • Results • significant higher concentrations of DNA recovered after automated extraction (p=0.001) • significant stronger Ct values for the automated solution (p=0.01)
Rapid & sensitive detection of large numbers of specimensApplication in Medical Diagnostics Detection of noroviruses in fecal specimensDr. J.A. Marshall at Victorian Infectious Diseases Reference Laboratory, Victoria (Australia) Experiment: total RNA extraction from fecal specimens + Analysis by multiplex RT PCR • Results • no significant difference between the manual and automation methods • automation methods required less material • automation methods process more samples in less time
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 21
SMARTQIAxtractor 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