Before you start

This topic lists the required hardware, software, AssayMAP Starter Kit, labware, and reagents for running the In-Solution Digestion protocol.

Hardware

Required hardware

 

 

The following figure and table show the components of the AssayMAP Bravo Platform, which is required for running the AssayMAP protocols.

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	Risers, 146 mm
7	Peltier Thermal Station with STC controller
8	Custom plate nest
9	Optional. Thermal plate insert for efficient heat transfer on the Peltier Thermal Station: • Red PCR Plate Insert (shown) • 96 Greiner U-Bottom Thermal Insert • 96 Greiner V-Bottom Thermal Insert • 96 AbGene U-Bottom Thermo Insert
10	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

•	Heated Incubator

•	Microplate sealer, such as the Agilent PlateLoc Thermal Microplate Sealer or equivalent

 

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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

 

 

 

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Labware

 

Use only the labware specified for each deck location. Using different labware or placing labware at unapproved deck locations can cause a collision resulting in equipment damage.

 

The In-Solution Digestion: Single Plate application does not require AssayMAP cartridges. All pipetting operations are handled in 96-well microplates using either pipette tips mounted on the AssayMAP probes or the bare AssayMAP probes.

The following figure shows the nine deck locations for labware.

Figure. Bravo deck locations (top view)

 

The following table provides a list of fully tested and approved labware types. The Sample plate type must be selected from the options available for deck location 4.

Table Labware options and deck locations

 

Labware	Manufacturer part number*	Deck location options
250-µL pipette tips (preloaded in 96AM Cartridge & Tip Seating Station, Agilent G5409-20025)	Agilent 19477-002	2
Lid, Universal**	Agilent 200858-100	4, 5, 6, 8, 9
96 Eppendorf 30129300, PCR, Full Skirt, PolyPro	Eppendorf 30129300	4, 5, 6, 8, 9
96 Bio-Rad PCR, Hard-Shell, Low-Profile, Full Skirt	Bio-Rad HSP‑9611	4, 5, 6, 8, 9
96 Greiner 652270, PCR, Full Skirt, PolyPro	Greiner 652270	4, 5, 6, 8, 9
96 Greiner 650201_U-Bottom, Clear PolyPro	Greiner 650201	4, 5, 6, 8, 9
96 Greiner 650207_U-Bottom, White PolyPro	Greiner 650207	4, 5, 6, 8, 9
96 Greiner 651201_V-Bottom, Clear PolyPro	Greiner 651201	4, 5, 6, 8, 9
96 Costar 3363, PP Conical Bottom	Corning Costar 3363	4, 5, 6, 8, 9
96 ABgene 1127, 1mL Deep Well, Square Well, Round Bottom	ABgene AB-1127	4, 5, 6, 8, 9
96 Eppendorf 96/500_V-bottom, Clear PolyPro	Eppendorf 96/500	4, 5, 6, 8, 9
96 Eppendorf 96/1000_U-bottom, Clear PolyPro	Eppendorf 96/1000	4, 5, 6, 8, 9
96 Waters 186005837, Clear PolyPro	Waters 186005837	4, 5, 6, 8, 9
12 Seahorse, Low Profile Reservoir, Natural PP	Agilent 201280-100	5, 6, 8, 9
8 Seahorse, Low Profile Reservoir, Natural PP	Agilent 201282-100	5, 6, 8, 9
Reservoir, Seahorse 201254-100, PP, 1 Well, pyramid bottom	Agilent 201254-100	5, 6, 8, 9
Reservoir, Axygen Scientific RES‑SW96-LP, 86mL pyramid bottom	Axygen Scientific RES‑SW96-LP	5, 6, 8, 9
96 V11 Manual Fill Reservoir	Agilent G5498B#049	5, 6, 8, 9
96 Red PCR Insert + Eppendorf 30129300, PCR, Full Skirt	Agilent insert (provided) and Eppendorf 30129300	4
96 Red PCR Insert + Bio-Rad PCR, Hard-Shell, Low-Profile, Full Skirt	Agilent insert (provided) and Bio-Rad HSP‑9611	4
96 Red PCR Insert + Greiner 652270, PCR, Full Skirt, PolyPro	Agilent insert (provided) and Greiner 652270	4
96 Greiner U-Bottom Thermal Insert + Greiner 650201_U-Bottom, Clear PolyPro	Agilent G5498B#126 and Greiner 650201	4
96 Greiner V-Bottom Thermal Insert + Greiner 651201_V-Bottom, Clear PolyPro	Agilent G5498B#126 and Greiner 651201	4
96 AbGene U-Bottom Thermal Insert + AbGene 1127, Square Well, Round Bottom	Agilent G5498B#127 and ABgene AB-1127	4
*For dimensionally equivalent alternatives and other labware details, see the Labware Reference Guide in the Literature Library page of the Protein Sample Prep Workbench. **Lidded plates are allowed at these deck locations, not the lid by itself. If you plan to use a lidded plate, ensure that you use only the approved lid type.

Table Labware options and recommended excess volume per well

 

Labware	Mfr part number	Recommended excess volume (µL) per well	Maximum capacity (µL) per well
96 Eppendorf 30129300, PCR, Full Skirt, PolyPro	Eppendorf 30129300	10	200
96 Bio-Rad PCR, Hard-Shell, Low-Profile, Full Skirt	Bio-Rad HSP-9611	10	200
96 Greiner 652270, PCR, Full Skirt, PolyPro	Greiner 652270	10	200
96 Greiner 650201_U-Bottom, Clear PolyPro	Greiner 650201	20	300
96 Greiner 650207_U-Bottom, White PolyPro	Greiner 650207	20	300
96 Greiner 651201_V-Bottom, Clear PolyPro	Greiner 651201	20	300
96 Costar 3363, PP Conical Bottom	Corning Costar 3363	20	300
96 ABgene 1127, 1mL Deep Well, Square Well, Round Bottom	ABgene AB-1127	30	1,000
96 Eppendorf 96/500_V-bottom, Clear PolyPro	Eppendorf 96/500	30	500
96 Eppendorf 96/1000_U-bottom, Clear PolyPro	Eppendorf 96/1000	30	1,000
96 Waters 186005837, Clear PolyPro	Waters 186005837	30	700
12 Seahorse, Low Profile Reservoir, Natural PP Lidded	Agilent 201280-100	3,000	6,500
8 Seahorse, Low Profile Reservoir, Natural PP Lidded	Agilent 201282-100	4,500	10,000
Reservoir, Seahorse 201254-100, PP, 1 Well, pyramid bottom	Agilent 201254-100	20,000	86,000
Reservoir, Axygen Scientific RES‑SW96-LP, 86mL pyramid bottom	Axygen Scientific RES‑SW96-LP	20,000	86,000
96 V11 Manual Fill Reservoir	Agilent G5498B#049	35,000	100,000

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

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Reagents

The types and amounts of reagents required to prepare for the In-Solution Digestion protocol depends on a combination of factors, including specific chemistry requirements, the number of samples to process, and volumes and concentrations of reagents necessary to conduct your digestion process. The Regent Volume Calculator for In-Solution Digestion calculates the reagent volumes required including the overages.

For more information, see Preparing the reagents.

 

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