Agilent gDNA Isolation Workstation
Application Bulletin

Summary• A flexible workstation providing
high-throughput gDNA isolation• Up to 10 microplates can be processed inone run without user intervention• Microplate processing time isapproximately 10 min per microplate(excluding incubation times)

Introduction
Efficient isolation of high-yield gDNA is
an important step in high-throughput
analysis of the genome. Isolating gDNA
can be accomplished by a number ofapproaches. Automated liquid-handlingworkstations can be used to isolate
gDNA in a reliable, repeatable, andcontamination-free manner. This application bulletin outlines a protocol for theInvitrogen ChargeSwitch EasyPlex gDNA
kit using the gDNA isolation workstationin which up to 10 microplates (96-well)
can be processed simultaneously.

+ + + ++ + + ++ + + ++ + + +B+L buffer
charges platesurfacepositively

Sample isadded to B+Lbuffer,DNAis released
from cells.

Negative
DNA backbone binds topositivitely
charged plate

gDNA Isolation Workstationconsisting of an Agilent
PlateLoc Thermal Microplate
Sealer (left), Agilent BenchCel
Microplate HandlingWorkstation (center), and an
Agilent Bravo Automated Liquid
Handling Platform (right).

System DescriptionMaterials

The workstation contains an Agilent Bravo
Automated Liquid Handling Platform, anAgilent BenchCel Microplate HandlingWorkstation, and an Agilent PlateLoc
Thermal Microplate Sealer. Microplates andtipboxes are delivered to the Bravo Platformby the BenchCel Workstation. The Bravo
gripper transports microplates and tipboxes
around the platform. Four reservoirs containing Binding and Lysis (B+L) buffer,
DNA sample solution, Wash-buffer 1 (W10),and Wash Buffer 2 (W12) are placed on theBravo deck. A recirculating reservoir is used
for liquid waste, and one deck location isdedicated for solid waste (tips, tipboxes).
All tips are exchanged after each step except
for the addition of the lysis buffer. After thewashing steps, the microplates are sealedwith the PlateLoc Sealer and stored in the
BenchCel Workstation for downstream use.

Component List
• Bravo Platform with gripper,
96LT disposable tip pipette head,
4 reservoirs, Liquid Waste Station• PlateLoc Sealer• BenchCel Workstation (Series R with6 stackers)• Agilent VWorks Automation
Control software

Washingsteps 1 and 2 remove
contaminants

Overview of the ChargeSwitch EasyPlex gDNA Isolation process

Isolated DNA isready fordownstream
applications

Labware List
• Microplate A: Invitrogen EasyPlex
96-well gDNA microplates• Tipbox A, B: Agilent Tips 96LT 200 µL
Reagent List
• Reservoir A: Binding andlysis (B+L) Buffer• Reservoir B: DNA sample• Reservoir C: Wash buffer W10• Reservoir D: Wash buffer W12

Instrument Layout

The Agilent Bravo deck (top view) withfour reagent reservoirs (locations 1-3,5), a solid waste station (location 6),
and a liquid waste station (location 4).

Conclusions

The Agilent BenchCel 6-stacker (top view) with microplates in stacker 1 and tipboxes instackers 4-6 (can store up to 50 tip boxes)

Protocol Workflow
1. Move tipbox A from BenchCel stacker
4 to Bravo location 7.
2. Move tipbox A from location 7 to 8.
3. Move microplate A from BenchCelstacker 1 to Bravo location 7.
4. Press on tips at location 9.
5. Aspirate 100 µL Lysis buffer fromreservoir A and dispense intomicroplate A.
6. Release tips at location 9.
7. Press on tips at location 8.
8. Aspirate 10 µL of sample from reservoir
B and dispense into microplate A.
9. Mix 5 times with tips.
10. Release tips at location 8.
11. Move tipbox from location 8 to 6.
12. Move microplate A from location 7 toBenchCel stacker 2.
13. Incubate 30 min.
14. Move microplate A from BenchCelstacker 2 to location 7.
15. Move microplate A from location 7 to 8.
16. Move tipbox A from BenchCel stacker
4 to location 7.
17. Press on tips at location 7.
18. Aspirate from microplate A, and empty
tips at liquid waste on location 4.
19. Release tips at location 7.
20. Move tipbox from location 8 to 6.
21. Move tipbox A from BenchCel stacker
4 to Bravo deck location 7.

22. Press on tips at location 7.
23. Aspirate 120 µL Wash buffer fromreservoir C at location 2 and dispense
into microplate A at location 8.
24. Mix two times with tips.
25. Aspirate from microplate A atlocation 8, and empty tips to liquid
waste at location 4.

The Agilent gDNA Isolation Workstation,utilizing the Invitrogen ChargeSwitchEasyPlex kit, provides the throughput, andwalk-away time necessary to keep up withthe demands of genomic applications. TheAgilent Bravo Automated Liquid Handling
Platform has the capacity for the necessary
reagents, plates, and tips in a space-saving
size. The integration of three Agilent
BenchCel stackers for tipboxes provides anuninterrupted supply of tips without userintervention. The Agilent PlateLoc Thermal
Microplate Sealer securely seals the plateafter processing, and is easily integrated
into the process workflow. The estimatedthroughput of the outlined protocol is 10 minutes for each plate processed (depending on
exact liquid handling procedures). Agilent
VWorks Automation Control software uses a
simple drag-and-drop method to create theevent-driven protocol for the entire process.
VWorks software includes error checking,error recovery, event reporting, and useraccess management.

26. Release tips at location 7.
27. Move tipbox from location 7 toWaste at 6.
28. Press on tips at location 7.
29. Aspirate 120 µL Wash buffer fromreservoir D at location 3 and dispense
into microplate A at location 8.
30. Mix two times with tips.
31. Aspirate from microplate A, empty toliquid waste at location 4.
32. Release tips at location 7.
33. Move tipbox from location 7 to waste
at 6.
34. Move microplate A from location 8 to 7.
35. Move microplate A from location 7
to PlateLoc Sealer.
36. Apply seal to microplate A.
37. Move microplate A to BenchCelstacker 3.

www.agilent.com/lifesciences/automation
This item is intended for Research Use Only.
Not for use in diagnostic procedures. Information,
descriptions, and specifications in this publication
are subject to change without notice.
Agilent Technologies shall not be liable for errors
contained herein or for incidental or consequential
damages in connection with the furnishing,performance, or use of this material.
Promega and Wizard are registered trademarks ofPromega Corporation.
Invitrogen and ChargeSwitch are registered trademarks of Invitrogen Corporation. EasyPlex is atrademark of Invitrogen Corporation.
© Agilent Technologies, Inc., 2009
Published in the U.S.A., February 26, 2009
Publication Number 5990-3549EN