eliminating bottlenecks with the agilent encore multispan ... · your workflow or do they…? a...

Eliminating Bottlenecks with the Agilent Encore Multispan Liquid Handler

Louis MurrayApplications Market Development & Software Product ManagerAgilent Automation Solutions

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.

A Few Thoughts on Bottlenecks & Automation

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…?

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

SOME EXAMPLES OF CHANGING BOTTLENECKS

…. AND SOME CHALLENGES WE STILL FACE

Miniaturization

96 well

384 well

1536 well

3456 well

Cell culturing for assays ...... Still quite laborious

Some improved consumables help with this bottleneck

GAME CHANGING TECHNOLOGIES

Sequencing, game changing technology…..

Bacterial genomes sequenced and publicly available.Source: www.genome.jp/kegg/

…..and moreGel Capillary

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

An Example of Moving Bottlenecks in the Next Generation Sequencing Workflows.

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

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

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

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

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

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

How Agilent addresses these bottlenecks

How Agilent is Approaching BottlenecksAutomate from Sample to Analysis

The Encore Multispan Liquid Handling System

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

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

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

Creating a Complete Walkaway Workflow

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

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%

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