eliminating bottlenecks with the agilent encore multispan ... · your workflow or do they…? a...
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Eliminating Bottlenecks with the Agilent Encore Multispan Liquid Handler
Louis MurrayApplications Market Development & Software Product ManagerAgilent Automation Solutions
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What is a bottleneck?
Definition: a phenomenon where the performance or capacity of an entire system is limited by a single or limited number of components or resources.
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A Few Thoughts on Bottlenecks & Automation
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Reasons to Automate in the Lab
Why Automate?1. Increase Throughput2. Increase Walkaway Time3. Increase Data Reproducibility
Both 1 & 2 remove bottlenecks from your workflow
Or do they…?
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A Consideration About Automation
Automation follows the laws of conservation:Like energy, bottlenecks are neither created nor destroyed. They are simply transferred from one area to another
• Automation never removes a bottleneck from your process, it simply moves it to another area in your lab
• Where it moves to will influence what new equipment you may need and where in the process it should be deployed
• Planning for this can help coordinate your automation and keep it running at maximum efficiency
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SOME EXAMPLES OF CHANGING BOTTLENECKS
…. AND SOME CHALLENGES WE STILL FACE
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Miniaturization
96 well
384 well
1536 well
3456 well
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Cell culturing for assays ...... Still quite laborious
Some improved consumables help with this bottleneck
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GAME CHANGING TECHNOLOGIES
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Sequencing, game changing technology…..
Bacterial genomes sequenced and publicly available.Source: www.genome.jp/kegg/
…..and moreGel Capillary
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Agilent RapidFire High-throughput MS SystemThe most efficient system for MS analysis
Faster Time to Results
LowerCost
• 8 –15 seconds per sample• >240 samples per hour
• Compatible with standard LIMS• No deviations to laboratory workflow• Data output format easily integrates
• 10x less solvent usage• Cartridge cost <10₵ per sample• >10x more efficient than LC/MS/MS
EasyOperation
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An Example of Moving Bottlenecks in the Next Generation Sequencing Workflows.
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NGS workflows, throughput and hands-on time
Agilent Bravo Liquid Handler drives throughput and hands-on time by automating the pipetting and streamlining the protocol with use of on-deck accessories and movement of on-deck labware by the gripper.
Addressing the first bottleneck
Hands‐On Timefor Manual Prep
Maximum Number of Samples
Processed/Day1Hands‐On Time
for Automated Prep
Maximum Number of Samples
Processed/Day
Library Prep 375 20 25² 96Pre‐Capture PCR 90 20 13 96
Hybridization Preparation 60 20 7 96
Capture 210 20 20 96Post‐Capture PCR 90 20 40 96
Total Minutes 82520 ‐ 40 per week
105192 per week
Total Hours 13.75 1.75
¹Assumes individual tube preps with an experienced user staggering sample processing²Library prep master mixes done once
Example showing throughput and hands‐on time while automating SureSelectXT on the BravoComparison is for automated v/s manual methods
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NGS AutomationMoving bottlenecks from library prep to sample normalization
Biological Sample
Purified DNA
Normalized Sample
Target Enriched Libraries
Pooled Sequencing Sample
Sequence Data
Analyzed Results
Encore Multispan + Bravo
Sequencing
Normalization
Pooling
Enrichment
Sequencing
Data Analysis
Bravo
NA Extraction
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1) Assess initial sample
concentrations
2) Create appropriate dilutions into TE
buffer
3) Confirm post normalization sample
concentrations
A Typical Nucleic Acid Normalization WorkflowWorkflow Summary
• Certain genomics workflows require samples to be initially “normalized” to a defined concentration and volume prior to further processing
• Example: NGS Library Prep Sample Requirements• HaloPlex: 45 uL at 5 ng/uL• SureSelect: 130 uL at 23 ng/uL
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A Typical Sample Normalization WorkflowNucleic Acid Normalization
Concentration Determination
Dilution Plate Creation
Add Stock Nucleic Acid
Sample
Post Normalization
Conc. Determination
Where do we start to automate?
Bulk Transfer
Low Volume Pipetting
Plate Reading
Plate to Plate
Transfer
Tube to Plate
Transfer
Plate Reading
Plate to Plate
Transfer
Tube to Plate
TransferPlate to
Plate Transfer
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A Typical Sample Normalization WorkflowNucleic Acid Normalization
Concentration Determination
Dilution Plate Creation
Add Stock Nucleic Acid
Sample
Post Normalization
Conc. Determination
Bulk Transfer Low Volume
Pipetting
Plate Reading
Plate to Plate
TransferTube to Plate
Transfer
Plate Reading
Plate to Plate
Transfer
Tube to Plate Transfer
Plate to Plate
Transfer
Where do we start to automate?• Liquid handling steps stand out as a major
contributor to the workflow
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A Typical Sample Normalization WorkflowNucleic Acid Normalization
Concentration Determination
Dilution Plate Creation
Add Stock Nucleic Acid
Sample
Post Normalization
Conc. Determination
Bulk Transfer Low Volume
Pipetting
Plate Reading
Plate to Plate
TransferTube to Plate
Transfer
Plate Reading
Plate to Plate
Transfer
Tube to Plate Transfer
Plate to Plate
Transfer
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How Agilent addresses these bottlenecks
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How Agilent is Approaching BottlenecksAutomate from Sample to Analysis
The Encore Multispan Liquid Handling System
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Encore Multispan Liquid Handling SystemAddress All Bottlenecks at Once
Flexible Multispan Pipetting• Pipettes twice as many samples with pipettors that can move
independently across 2 dimensions• A variety of on deck accessories such as plate shakers and temperature
control compliments your workflow, allowing for significantly higher walkaway time
Built-In Robotic Arm• Robust management of labware both on and off deck• Integrate up and downstream processes and create a complete
walkaway workflow• Have the flexibility to rearrange your automation with virtually no
down time
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Individual Pipettor Performance From 0.3uL to 200uL
0
1
2
3
4
5
6
7
0.3 0.4 0.5 0.6 0.8 1 10 25 30 50 55 200
10µl tip (DMSO)
10 µL tip (aq.)
30 µL tip (aq.)
70 µL tip (aq.)
250 µL tip (aq.)
%CV
Volume (uL)
Instrument Specification
• Instrument Specification= 5%CV• n=8 for each data point
• Tested w/ Artel MVS• Full aspirate/dispense with leading airgap
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A Complete Walkaway Solution for Sample Normalization
Benefits
• User definable target concentration and volume• Enables inclusion of quantification reagent (e.g. Pico Green)• Hands-off• Scalable• Flexible sample input format (tubes or plates)• Final concentration verification
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Creating a Complete Walkaway Workflow
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A Typical Sample Normalization Workflow (Revisited)
Concentration Determination
Dilution Plate Creation
Add Stock Nucleic Acid
Sample
Post Normalization
Conc. Determination
Bulk Transfer Low Volume
Pipetting
Plate Reading
Plate to Plate
TransferTube to Plate
Transfer
Plate Reading
Plate to Plate
Transfer
Tube to Plate Transfer
Plate to Plate
Transfer
• The entire sample normalization workflow is now completely walkaway
• Results are not be affected by operator variability
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Nucleic Acid NormalizationOur results
• 96 samples normalized down to 20 ng/uLfrom their determined initial concentration
• Encore Multispan was able to normalize samples with concentrations as high as 44x with high precision
• All 96 samples had initial and final concentrations determined with Picogreenreagent
• Results Statistics• Average Conc. (ng/uL) 22.29• Overall CV 7.4%
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Thank You!
For more information, please visit:http://www.chem.agilent.com/en-US/products-services/Instruments-
Systems/Automation-Solutions/Encore-Multispan-Liquid-Handling-System