Figure 1. Laboratory set up. All automated experimental set ups were carried out on a Gilson Pipetmax housed in a sterility hood, alongside a Quant Studio 5 qPCR thermocycler, collectively controlled via the Antha software.

Figure 2. The Antha Workflow Editor. Antha provides a sand box environment, the Workflow Editor, for rapid liquid handling workflow development utilising the Standard Element Set.

An example Antha qPCR workflow developed to address Cambridge Consultants’ specific assay requirements, incorporating sample re-suspension, splitting of positive and negative control samples for storage and future re-use, serial dilution of qPCR standards, aliquoting of qPCR master mix and final qPCR reaction set up.

 

While this is a well-known technique, the Cambridge Consultants team had some specific needs, requiring the flexibility to rapidly iterate and optimise, that made it more challenging to develop this workflow through the vendor’s software platform. In particular:

  • Consistently high throughput to run many thousands of samples during the project.

  • Fast execution speed to process up to a thousand unique samples per day.

  • The ability to add custom sample handling steps upstream of qPCR reaction set up (Figure 2).

  • Robust sample traceability to manage the data and its provenance (Figure 3).

  • Excellent reproducibility to allow optimisation of underlying process (Figure 4).

  • Minimising execution costs given the scale of testing (reduced FTE time, Figure 6).

The results of the experiments (Figure 5) are used as part of formal acceptance testing therefore liquid handling reliability is essential (especially cross-contamination control) along with clear traceability where both the liquid handling platform and analytic device are under the control of a single software solution.