Single Liquid Addition v1.1

The Single Liquid Addition Utility is specifically designed to adjust the pH of samples after the samples have been digested using the In-Solution Digestion: Multi-Plate application. This utility enables you to rapidly adjust buffer composition or add a component to up to four sample plates by transferring liquid from a common reagent plate, to up to four independent sample plates. An optional stringent syringe wash step is included to prevent carry over.

Before you start

Make sure you:

•	Use Greiner 650201 or 650207 U-Bottom Plates for the sample plates.

•	Place up to four sample plates on the Bravo deck, at the following locations:

–	5—Sample plate 1

–	6—Sample plate 2 (optional)

–	8—Sample plate 3 (optional)

–	9—Sample plate 4 (optional)

•	Prepare a reagent plate that contains the solution that you want to add to the sample plates.

–	For the list of approved labware, see the labware list in the C. Labware Table area of the Single Liquid Addition utility. See Accessing the Single Liquid Addition utility.

–	Add the recommended excess volume in the reagent plate. The extra volume requirements vary by plate type.

You can find the labware-specific values in the Labware Reference Guide, which is in the Literature Library page of the Protein Sample Prep Workbench.

•	Place the reagent plate at deck location 4.

•	Prepare the Syringe Wash Buffer. For suggested solutions to use, see Preparing the reagents in the In‑Solution Digestion: Multi-Plate v1.1 User Guide.

•	Ensure that you have four Greiner 650207 White U-Bottom plates for the wash plates. You must fill the plates with the Syringe Wash Buffer, and then run the Plate Stacking protocol to stack the wash plates at Bravo deck location 2.

Accessing the Single Liquid Addition utility

To access the Single Liquid Addition utility:

Open the Utility Library, locate the Single Liquid Addition utility, and then click Utility.

 

The following figure shows the Single Liquid Addition utility interface.

 

 

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Running the Plate Stacking protocol

The Plate Stacking protocol stacks the wash plates properly at Bravo deck location 2 to prevent collision during the Single Liquid Addition protocol run.

 

The Single Liquid Addition protocol requires the stack of four wash plates, even if you have less than four sample plates. Ensure that the wash plates are Greiner 650207 White U-Bottom.

 

Before running the Plate Stacking protocol, prepare four Greiner 650207 White U‑Bottom plates as follows:

•	Fill the plates with 300 µL Syringe Wash Buffer. The plates will serve as wash plates that are used for cleaning the syringes between liquid transfers.

•	Make sure the filled wells correspond to the location of the samples in the sample plates.

To prepare the Wash Plates:

1	In the Single Liquid Addition utility, click Run Plate Stacking. The protocol starts.

2	Follow the on-screen instructions for placing the wash plates at the appropriate locations on the AssayMAP Bravo deck.

3	Check the Status box in the upper right corner of the form. When the Protocol Complete message is displayed, click OK.

 

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Running the Single Liquid Addition utility

To run the utility:

1	In the A. Application Settings area, set the following properties for the run:

 

Property	Description
Number of Initial Syringe Washes	The number of cycles for rinsing syringes with deionized water at the beginning of the run. Range: 0–10 Default: 3
Number of Sample Plates	The number of sample plates (1-4) to which a liquid will be added in this utility. Place only the number of sample plates used on the deck. Positions 2-4 are optional and need not be filled. Range: 1–4 Default: 1
Initial Volume in Sample Plate	The initial volume of the samples. All plates must have the same sample volume. Range: 0–250 Default: 10
Volume to Add to Sample Plate	The volume of solution being added to the samples. This may be limited by the labware. Range: 0–250 Default: 50
Number of Mixing Cycles	The number of mixing cycles that will be used to homogenize the sample liquid and the liquid that is added. Range: 0–20 Default: 10
Conduct Stringent Syringe Wash	The option to wash the syringe with a solution in the wash plate between transfers. Select the check box to perform the wash. Clear the check box to skip the wash. Default: 
Number of Syringe Washes between Transfers	The number of Syringe Washes to be performed at the Wash Station, before adding liquid to the next sample plate. Range: 0–10 Default: 5
Number of Final Syringe Washes	The number of times to rinse syringes with deionized water. Range: 0–10 Default: 3

2	In the Labware Table, select that labware you are using at deck location 4.

 

Incorrect labware selections can cause a hardware collision, resulting in equipment damage. Ensure that the selections in the Labware Table exactly match the physical labware present on the Bravo deck.

 

3	Place the labware on the AssayMAP Bravo deck locations, as shown in the Deck Layout area.

4	Ensure that the labware are properly seated in the platepads.

 

Improperly seated labware can cause a hardware collision, resulting in equipment damage. Ensure that all labware are properly seated within the alignment features of their respective platepads.

 

5	Click Run Liquid Addition to start the protocol run.

To monitor the progress of the run, check the Status box in the upper right corner of the form.

When the run is finished, remove the labware from the deck.

 

To stop a run in an emergency, use the emergency stop pendant.

 

To pause the run, click Pause. The task currently in progress finishes before the protocol pauses. The Scheduler Paused dialog box opens. For details, see Emergency stops and pauses.

To troubleshoot errors, see the Error Recovery Guide and the Bravo Platform User Guide in the Literature Library page of the Protein Sample Prep Workbench.

 

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