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GenomeLab™ Gene Expression Suite: Using the Biomek® 3000 Laboratory Automation Workstation to Perform Quantitation and Normalization of Nucleic Acid Samples

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Amy Yoder, Matthew Cu, Kelly Marshall, M.S. Laura Pajak, Ph.D. Beckman Coulter, Inc.

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Figure 1. Flow Chart for Quantitation and Normalization Method using the Biomek 3000 Workstation

GenomeLab GeXP Genetic Analysis System using the Biomek 3000 Laboratory Automation Workstation. Quantitation and Normalization is the second method in the suite and is the focus of this poster. The Quantitation and Normalization Method is a multipurpose utility designed to quantitate source nucleic acids in a 96-well plate format and normalize the samples to user-defined volumes and concentrations. The method has default parameters for downstream GeXP applications and the user interface allows for customization of many parameters thus permitting the method to be used for applications beyond GeXP sample preparation. The options include nucleic acid type, number of samples, and the choice of quantitation via absorbance at 260 nm or fluorescence using Invitrogen’s Quant-iT* reagents. Standard curve parameters, plate type, and dilutions can also be defined based on the amount and expected concentration of the samples. An integrated calculations template allows for conversion of quantitation data from the DTX 880 Multimode Plate Reader to values to be used by the normalization method. The template records the user defined variables in the Biomek method and allows users to enter their desired normalization parameters after the quantitation calculations are complete. Users can decide how to handle out-of-range samples by either excluding them from further processing or transferring the wells with low concentrations. The method normalizes all wells by volume and concentration with low coefficient of variation.

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Additional Information

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Application System Requirements

Hardware Options

Source	Part No.	Quantity	Description
Beckman Coulter, Inc.	986120	1	Biomek 3000 Laboratory Automation Workstation 50/60 Hz, 100-240 V
Beckman Coulter, Inc.	987921	1	DTX880 Multimode Reader
Beckman Coulter, Inc.	986146	1	Biomek 3000 MP200 Multi-Tip Pipette Tool: Capacity 5-200mL
Beckman Coulter, Inc.	987368	1	Biomek 3000 P200L Singe-Tip Pipette Tool: Capacity 5-200mL
Beckman Coulter, Inc.	987371	1	Biomek 3000 Gripper Tool
Beckman Coulter, Inc.	609641	1	Gripper Tool Rack

Software Options

Source	Part No.	Quantity	Description
Beckman Coulter, Inc.	719349	1	Biomek Software

Consumables and Reagents

Source	Part No.	Quantity	Description
Beckman Coulter, Inc.	717252	1	Biomek AP96 P250 Pipette Tips, presterile
Beckman Coulter, Inc.	717255	2	Biomek AP96 P20 Pipette Tips, presterile
Beckman Coulter, Inc.	609801	2	96-Well Sample Microtiter V-Bottom Thermocycler-Compatible Polypropylene Plate
Beckman Coulter, Inc.	373661	1	24-Position Tube Rack
Beckman Coulter, Inc.	373696	1	Insert, 11mm, White, for 1.5 mL Microfuge Tubes
Beckman Coulter, Inc.	356090	3	Polypropylene Tubes with Snap-On Caps, 1.5 mL
Beckman Coulter, Inc.	372795	1	Modular Reservoir Frame

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* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffmann-LaRoche, Ltd.

For Research Use Only; not for use in diagnostic procedures.

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