Preparing the samples
To minimize evaporation, prepare the samples immediately before running the Affinity Purification protocol, or keep the plates lidded until the run begins.When preparing the samples, you must:
• Choose a sample plate with sufficient capacity to hold the total reaction volume, including the sample volume and the volume for all the completed addition steps in the run.
• Transfer the samples to the microplate you want to use for the protocol run.
Planning the microplate setup
Before transferring the samples, you should plan the layout of the samples in the microplate. Consider the following:
• You can process 1 to 96 samples in parallel. The samples should be arranged in contiguous columns. The position of the samples in the microplate dictates the positions of the buffer solutions in the microplates and reservoirs.
• If you have fewer than 96 samples, make sure the samples occupy full columns in the microplate.
The default protocol settings assume that samples will be arranged in multiples of 8 in a column-based configuration.
If using pipette tips for transfer, the AssayMAP Bravo Platform applies differential pressure to seat pipette tips based on the number of full columns of pipette tips. To achieve proper pipette tip seating, entire columns must be used.
Transferring the samples to the microplate
A small volume excess is required in all labware types to ensure proper volume transfer. An excess (overage) volume ensures that a microplate well does not fully deplete, which would result in aspiration of air into the syringes or the pipette tips, compromising performance.The Reagent Volume Calculator shows the recommended overage for the labware types being used and automatically includes recommended overages in the volume it recommends per well.
Labware-specific overage recommendations are also presented in the Labware Reference Guide, which you can find in the Literature Library page of the Protein Sample Prep Workbench. More or less overage can be used depending on the solution and the length of the run but the recommended overages are fine for most standard runs.
To transfer the samples to the microplate:
1 Run the Reagent Transfer utility or Reformatting utility to transfer the samples. For instructions, see one of the following:
2 If necessary, centrifuge the sample labware to remove bubbles.