Preparing the samples

 

To minimize evaporation, prepare the samples immediately before run time or keep them covered until you run the protocol.

 

The In-Solution Digestion application can accommodate a wide range of protein concentrations, which is largely dependent on downstream needs. Viscous samples may require dilution.

When preparing the samples, you must:

•	Remove macromolecular particulates before the samples are loaded onto AssayMAP cartridges.

•	Determine the volume of samples.

•	Transfer the samples to the microplate you want to use for the protocol run.

Removing macromolecular particulates

Make sure the samples are free of macromolecular particulates, such as large protein aggregates and cellular debris. If the samples have passed through a chromatography column, they should be free of particles and can be loaded on to an AssayMAP cartridge. Samples that might contain particles should be filtered through a 0.22-µm filter before loading on an AssayMAP cartridge.

 

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Adjusting the sample composition

The concentration of denaturant in the sample should be entered into the In-Solution Digestion Reagent Volume Calculator. See About the Reagent Volume Calculator for In-Solution Digestion.

 

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Determining the volume of sample to load

The acceptable volume of the sample is highly dependent on the digestion conditions. Use the In-Solution Digestion Reagent Volume Calculator to help determine sample volume. See About the Reagent Volume Calculator for In-Solution Digestion.

 

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Transferring the samples to labware

Setting up the sample microplates

Before transferring the samples, you should plan the layout of the samples in the microplate. Consider the following:

•	You can process 8 to 384 samples in parallel, and up to four sample plates.

•	The volume of samples is limited by the labware you use and the digestion reaction conditions. See About the Reagent Volume Calculator for In-Solution Digestion. The total volume should also include any volume of liquids you intend to add post digestion.

Transferring the samples to the microplate

You can transfer the samples to the microplate that is supplied with the AssayMAP Starter Kit. See Labware and starter kits.

To transfer the samples to the microplate:

1	If you have multiple sample plates, run the Single Liquid Addition utility to transfer the samples. For instructions, see Single Liquid Addition v1.1.

If you have only one sample plate, run the Reagent Transfer utility to transfer the samples. For instructions, see Reagent Transfer v2.1.

2	If necessary, centrifuge the sample labware to remove bubbles.

 

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