Sample & Assay Technologies - 1 - smart compact affordable Brant Bassam PhD QIAxtractor Andrea...
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Transcript of Sample & Assay Technologies - 1 - smart compact affordable Brant Bassam PhD QIAxtractor Andrea...
Sample & Assay Technologies- 1 -
smart compact affordable
Brant Bassam PhD
QIAxtractor
Andrea TesorieroApplication Specialist
Sample & Assay Technologies- 2 -
QIAxtractor is a seamless integration to your workflow
post PCR analysis
real timePCR or RT PCR
PCR setup nucleic acid
extractionSample
preparation
PCR &analysis
PCRsetup
enzymaticdigestion
post purification
QC
enzymaticdigestion
aliquoting DNA
extractionSample
preparation
Sample & Assay Technologies- 3 -
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
Sample & Assay Technologies- 4 -4
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
Sample & Assay Technologies- 5 -5
5 Minimize cross contamination risk5 Reduce human error5 Reduce costs5 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
5 Increase safetyMinimum exposure to dangerous chemicals
5 Versatility and choiceDX liquid or tissue reagent,Vx reagent Wide range of protocols for DNA, RNA and viral nucleic acidsAssociated plasticware
5 Maintain your sanity (pain threshold is about 25 extractions per week!)
Why automate?
Sample & Assay Technologies- 6 -6
Sample types successfully extracted
5Blood -whole, fresh, frozen, buffycoat,Human, Mouse, Turtle, chickens, sheep Bovine Serum/Plasma, -HIV, Hep B , Hep C, CMV
5Urine Chlamydia and N. Gonorrhoea
5Swabs HSV1 and 2
5Solid Tissue Mouse Tails, Prawn Tissue, Bird Quill Pulp
5Nasal Aspirates MPV (a single stranded RNA Virus)
5Cultured cells
5Micro Organisms Bacteria (Gram Pos. and Gram Neg.) and Fungi
5Plants Fruit, bark, leaves and stems
5Faeces Norris Virus
5Forensics cigarette butts, food scraps, finger prints, hair, fabric, FTA cards
Sample & Assay Technologies- 7 -7
5Easy to use system
5Low cost possible cost per extraction
5Hands-free, walk-away operation
5High speed + high throughput (96 well)
5Consolidated chemistryminimum reagents and kit types work for all sample types
5Complete integrated system solutionautomation, reagents, consumables, protocols, options
QIAxtractor
Sample & Assay Technologies- 8 -8
490 mm
390
mm
clo
sed
770
mm
ope
n
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
1122 8-Channel
pipetting head
Sample & Assay Technologies- 9 -9
19 kg
520
mm
495 mm400
mm
Under BenchVacuum/Waste Station
Waste bottle4 L sanitary quick disconnect bottle
Bottle full sensor
Vacuum sensorSoftware 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
Sample & Assay Technologies- 10 -10
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
Sample & Assay Technologies- 11 -11
CorProtocols™ for different applications
Available for web download
Each comprises:
PDF Protocol (Instructions for Use)
X-tractor Gene Run File
Sample & Assay Technologies- 12 -12
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)
Sample & Assay Technologies- 13 -13
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
Sample & Assay Technologies- 14 -14
Barcode Reader Compatible
Sample names and information can also be imported or exported
Sample & Assay Technologies- 15 -15
Pre- and Post-Run Reports
5HTML report (universal format)
5Provides details of a run
5Documentation & Traceability
(e.g. sample, batch and lot identifiers)
5Can be directly: Printed Saved E-mailed Exported to Word
Sample & Assay Technologies- 16 -16
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
Sample & Assay Technologies- 17 -17
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 Tip96 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
Sample & Assay Technologies- 18 -
QIAxtractor minimizes cross-contamination riskReal-time amplification analysis of all samples
Real-Time AnalysisReal-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-contaminationCycle
5 10 15 20 25 30 35 40
Nor
m. F
luor
o.
0.25
0.20
0.15
0.10
0.05
0.00
Threshold
DX Reagent Verification Run■ 48 Positive samples
■ 48 NTC samples
Maximum CT 25.04
Minimum CT 24.19
Average CT 24.59
CT Std. Dev. 0.18
Sample & Assay Technologies- 19 -
QIAxtractor optimizes nucleic acid extraction yieldApplication 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 concentrationfrom 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 qualitybetween manual and automated DNA extraction methods by real time analysis
Viral detection of EHV-1 horsesDr. N. Pusterla at University of California, School of Veterinary Medicine
Sample & Assay Technologies- 20 -
Rapid & sensitive detection of large numbers of specimensApplication 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 2h - -
Time for 96 specimens
- - 2h30 1h30
Time Operator was actually busy
2h 2h 25 min 15 min
Experiment: total RNA extraction from fecal specimens + Analysis by multiplex RT PCR
Detection of noroviruses in fecal specimensDr. 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
Sample & Assay Technologies- 21 -21
Benefits
5Consistency & Workflow Reduce human error and free-up technician time
5Standardisation Between labs and runs
5Documentation Pre- and Post-Run reports
5Safety 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
Sample & Assay Technologies- 22 -
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
Sample & Assay Technologies- 23 -
5Gracias por su atención