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


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


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


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 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


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


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


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


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


CorProtocols™ for different applications

Available for web download

Each comprises:

PDF Protocol (Instructions for Use)

X-tractor Gene Run File


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)


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


Barcode Reader Compatible

Sample names and information can also be imported or exported


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


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


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


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


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


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


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


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


5Gracias por su atención

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

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