To create a pH gradient you combine 2 buffers which are of the same type but are at different
stock pH values.
In order to do this, you first need to set up a 'pH System'.
- Click on the 'pH Systems' tab in the Components section, then click ‘Add New’.
- To change the name, maximum pH range, stock concentration and pKa values, click on ‘Edit…’
- Click on ‘Edit’. Enter the name of a buffer (only buffer ingredients are available), such as TRIS.
- The pH system now appears as a Component on the 'pH Systems' tab.
Using a pH System to create a gradient is very similar to standard concentration gradients. First
select the desired region. Select the ‘pH gradients’ tab and click ‘+ add pH system’.
Then select the buffer system from the drop-down menu. Choose the gradient direction to be
applied to the region and enter the constant concentration which the buffer mix should be at
throughout. The pH values chosen cannot exceed the range of the pH system.
Edit the pH gradient range and concentration in the panel above the plate map grid.
Note: although regions can overlap, pH gradients cannot: each well can contain only one pH value.