Before you start

This topic lists the required hardware, software, labware, and solutions for running the Normalization protocol.

Hardware

Required hardware

 

 

The AssayMAP Bravo Platform is required for running the AssayMAP protocols. The following figure and table describe the platform components.

Figure. AssayMAP Bravo Platform components

 

 

Item	Required hardware
1	Gripper upgrade
2	Bravo 96AM Head
3	96AM Wash Station
4	Pump Module 2.0 and two carboys
5	96AM Cartridge & Tip Seating Station
6	Bravo Risers, 14.6 cm
7	Peltier Thermal Station with STC controller
8	Plate Risers, 2.84 cm (two)
9	Orbital Shaking Station with Control Unit

 

 

 

Optional equipment

You might need the following when preparing the samples and reagents:

•	Microplate centrifuge, such as the Agilent Microplate Centrifuge or equivalent

•	Microplate sealer, such as the PlateLoc Sealer or equivalent

 

Return to top

Software

 

 

The following table lists the minimum software requirements:

 

Software	Version
Agilent VWorks Automation Control	13.1.5
Agilent Protein Sample Prep Workbench	3.2
Microsoft Excel Required for the reagent volume calculators and method setup tools.	2016

 

 

 

Return to top

Labware

The Normalization protocol requires the following labware:

•	Full tip box of 96 250-µL pipette tips, Agilent 19477-002

 

Using different pipette tips, such as filtered tips, will result in equipment damage. Ensure that you use the standard 250-µL pipette tip (Agilent 19477-002).

 

•	Labware for the Diluent reservoir, initial Sample plate, and Normalized sample plate

The following tables list the labware options, the corresponding deck locations, and the excess volume recommendations for each option.

Table Labware options for the Diluent reservoir and the corresponding excess volume recommendations

 

Labware	Manufacturer part number	Deck location	Excess volume (µL) per column or reservoir
12 Column, Low Profile Reservoir, Natural PP	Agilent 201280-100	4	3,000
8 Row, Low Profile Reservoir, Natural PP	Agilent 201282-100	4	4,500
96 V11 Manual Fill Reservoir	Agilent G5498B#049	4	20,000
Reservoir, Axygen Scientific RES‑SW96-LP, 86mL pyramid bottom	Axygen Scientific RES‑SW96-LP	4	20,000

Table Labware options for Initial Sample plate (deck location 2) and Normalized plate (deck location 6)

 

Labware	Manufacturer part number	Deck location	Unusable volume (µL) per well
96 ABgene 1127, 1mL Deep Well, Square Well, Round Bottom	ABgene AB-1127	2, 6	30
96 Eppendorf 30129300, PCR, Full Skirt, PolyPro	Eppendorf 30129300	2, 6	10
96 Greiner 652270, PCR, Full Skirt, PolyPro	Greiner 652270	2, 6	10
96 Greiner 650201_U-Bottom, Clear PolyPro	Greiner 650201	2, 6	20
96 Greiner 650207_U-Bottom, White PolyPro	Greiner 650207	2, 6	20
96 Greiner 651201_V-Bottom, Clear PolyPro	Greiner 651201	2, 6	20
96 Costar 3363, PP Conical Bottom	Corning Costar 3363	2, 6	20
96 Greiner 675801, Half Area, Flat-Bottom, UV Star	Greiner 675801	2, 6	25

The unusable volume (also known as dead volume) is the volume that cannot be reliably aspirated from the microplate well due to pipetting limitations that arise from differences in well-bottom geometry. The following table provides some general guidelines on how different microplate geometries can affect this value.

Table Guidelines on how microplate geometry can affect volume requirements

 

Well-bottom geometry	Examples	Unusable volume expectations
Flat bottom	Greiner 675801	Generally, the worst plates for minimizing dead volume. Liquids are attracted to the intersection of the plate bottom and side walls.
Round (U) bottom	Greiner 650201, Greiner 650207	Generally, good options for minimizing dead volume. A round bottom forces liquid to collect in the center of the well at a medium rate.
V-bottom and PCR	• V-Bottom: Greiner 651201, Corning Costar 3363 • PCR Plate: Eppendorf 30129300, Greiner 652270	Generally, the best plates for minimizing dead volume. Steep side walls force liquid to collect in the center of the well, making it easy to access.

For dimensionally equivalent alternatives and other labware details, see the Labware Reference Guide in the Literature Library page of the Protein Sample Prep Workbench.

Return to top

Solutions

The Normalization utility requires the following solutions:

•	Diluent to adjust the sample concentration

•	Purified water for the wash station reservoir (carboy)

 

Return to top