Preparing the diluent and samples

 

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

 

Note: If you wish to access the diluent and serial dilution plate preparation instructions at a later date, you can do so by using Microsoft Excel to open the saved method file (.csv).

To prepare the diluent reservoir plate:

Follow the instructions in Step 7) Prepare serial dilution plates, which will indicate the diluent volume (µL) necessary for each dilution series replicate and the well locations where this diluent volume should be placed.

In addition to the diluent volume required to generate the serial dilution curve, some volume overage is required to account for the dead volume in the plate well. For recommended overage volumes and well capacities, see the Labware Reference Guide (workbench Literature Library page).

•	For 96-well plates, this volume calculation is straightforward in that it is the volume required plus the recommended overage for the well.

•	For plates where multiple wells on a 96-well plate map are combined in a common well, the situation can be more complex.

–

If only a single serial-dilution series will draw from the common well (e.g., using an 8-channel reservoir where the serial dilutions go from left to right) the situation is the same as in a 96-well plate where the volume required in the common well is the volume indicated in step 7 plus the required overage for the common well.

–	If more than 1 serial dilution will draw from a common well (e.g., using an 8‑channel reservoir where the serial dilutions are going from top to bottom and you have more than one replicate) you would add all the required volumes indicated in step 7 plus the required overage for the common well.

If necessary, use the Reagent Transfer utility to transfer the diluent into the appropriate labware type. For instructions, see Reagent Transfer v2.1.

To prepare the serial dilution plate:

Follow the instructions in Step 7) Prepare serial dilution plates, which will indicate the volume (µL) and concentration of the first dilution for each dilution series replicate and the well locations where this starting volume should be placed.

Ensure that the samples in the serial dilution plate match the volume and well positions specified in the method created using the Method Setup Tool.

If necessary, use the Reformatting utility to transfer the samples into the appropriate labware type. For instructions, see the Utilities: Reformatting v2.0 User Guide.

 

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