A miniaturized purification workflow can include the following elements:

They tell Synthace:

Which resins, samples and buffers you want to use
Where the resins are located on your robocolumns plate
When and how the liquid handler will perform a chromatography stage

Synthace uses the elements to generate a set of instructions for your device to follow.

A miniaturized purification workflow requires at least three elements from the RoboColumns set:

Define Resin Types and Data to Resin Types;
Define RoboColumns Plate; and
Run Chromatography Stage or Run Gradient Chromatography Stage.

Combine them with elements from the Core Elements set to build a complete miniaturized purification workflow.

Commonly used elements from the Core Elements set include:

Define Liquids and Plates. To learn more, click here.
Append. To learn more, click here.
Run Plate Reader.

Define Resin Types

This element tells Synthace which resins you have.

Use it to define the name of each resin, and any additional metadata (such as manufacturer name, batch number, etc.) that you want to add.

Data To Resin Types

This element is similar to the Define Resin Types element, but it imports the information from a spreadsheet.

If you want to add lots of information about the resins, this is the quicker option. You can upload from an external software or enter the information directly in Input Data (Table).

Define RoboColumn Plate

This element tells Synthace about the robocolumn array plate.

Use it to define:

How many robocolumns you have
Where they are located on the plate
Which resin is in each robocolumn
How much resin is in each robocolumn; and
The robocolumns names

Note: To save time, Synthace generates a name for each robocolumn.

Synthace needs to know this so that it instructs the liquid handler to dispense the samples and buffers into the correct wells.

Run Chromatography Stage

This element instructs the liquid handler to perform a chromatography stage. The element has three distinct settings.

Stage Setting

The stage setting allows you to specify the name of the chromatography stage. Synthace has pre-populated names you can select from, or you can enter your own.

Note: Each stage name must be unique. If you run the same stage multiple times, we recommend you number the name of each stage (ex. Equilibrate 1, Equilibrate 2, etc.).

Chromatography Settings

The chromatography settings allow you to define parameters for liquid loading of the chromatography stage.

Click on Configure to bring up the following window.

For each robocolumn, use this to define:

The liquid that you want the liquid handler to dispense into the robocolumn;
The total load amount and a way in which that volume should be fractionated (fractionating of the total load volume can be described in volumes or time to collect one fraction);
The volume that you want each fraction to be, expressed in column volumes or ul;
Whether to collect or send the fractions to the waste tray;
The residence time, volumetric flow rate or flow velocity at which you want the liquid handler to dispense the liquid; and
The liquid handling policy that you want the liquid handler to use when it aspirates liquid from its input plate.

Alternatively, the total load amount can be expressed in mass.

Tip: You only need to define one of either residence time, volumetric flow rate or flow velocity. Synthace uses the value that you specify and the volume of resin in each robocolumn to calculate the values of the other parameters.

If you wish to add a wait time between a dispense into a robocolumn and the next liquid handling action, use Post-Dispense Wait Time Overrides.

Note: currently, loading by mass is not available in Templates/Forms.

Collection Settings

The collection settings allow you to describe what you would like to do with your flow.

If you wish for the flow through to pass to the waste tray, do not toggle on Collect Flow Through.

If you wish to collect your fractions, toggle on Collect Flow Through.

Here you can specify:

To align your collected fractions at the start or end.
- Note: This is typically only used when using different flow rates for the same chromatography step. Click on the question mark next to Collection Alignment to learn more.
The plate in which you want to collect your fractions, the collection plate name and plate type.
Whether or not the collection plate has a lid.
Whether or not to aliquot a volume of the liquid into a well for blank correction.
- Note: This will cause the liquid to be plated directly on the collection plate without passing through robocolumns.
- Blanks can now be expressed in CV or ul;
- Alternative blanks can be specified for a given Run Chromatography Stage (eg. wash buffer blank for the load stage). Simply provide the blank volume and the name of the liquid you wish to blank correct against in the Blank collection Settings.

Run Gradient Chromatography

This element is similar to the Run Chromatography Stage element, but it does not instruct the liquid handler to dispense one buffer into each robocolumn.

Instead, it instructs the liquid handler to create a series of mixtures from Buffer A and Buffer B (one per fraction that you want to collect) and dispenses them into the robocolumns. To define this, click on Configure within Load Parameters.

Use this element to control the percentage of Buffer B that you want the liquid handler to add to the first and last mixtures in the series.

Synthace uses these percentages to calculate the appropriate ratio of Buffer A to Buffer B for the intermediate mixtures.

In this example, we are collecting six fractions from a robocolumn. By setting the Buffer B Start Percentages to 20% and the Buffer B End Percentages to 70%, Synthace will instruct the liquid to create six mixtures. They will have the following ratios of Buffer A to Buffer B.

80% : 20%
70% : 30%
60% : 40%
50% : 50%
40% : 60%
30% : 70%

All other settings on Run Gradient Chromatography Stage are the same as on Run Chromatography Stage, apart from loading by mass.