BioLT

Worked Example

Pipetting Overage Planning Example

Worked example for deciding how much extra master mix to prepare when small dead volumes and repeat dispensing are expected.

Scenario

You need enough reaction mix for 24 wells, but the exact calculated volume leaves no margin for tip retention, tube dead volume, or small transfer losses.

The example shows how to choose overage as a practical bench decision rather than a random percentage.

Inputs to confirm

Number of reactions and final volume per reaction.

Whether the mix will be dispensed from a tube, strip, or reservoir.

Whether the limiting reagent can tolerate extra prepared volume.

How to use the result

Calculate the exact required total, then add a defined overage such as 10 percent or one extra reaction depending on scale.

Record the overage rule so the next person understands why the prepared volume is larger than the mathematical minimum.

Common failure points

Preparing exactly the required volume and running short in the final wells.

Adding too much overage when an expensive enzyme or rare sample is limiting.

Changing overage between plates without recording the reason.