Accuracy Matters When Quantitative, Manually-pipetted Assays Graduate to Automation

A Story in Diagnosing and Troubleshooting


It is often the case that assays are initially performed on the benchtop  using handheld  pipettes  before they graduate, or transfer,  to an automated liquid handler.  Automating a manual method may take time and patience,  but automation will help lower costs, increase  throughput, and potentially avoid errors associated with a manual method. During the transfer  process,  however,  the manual assay should be directly compared to the automated assay for consistencies in pipetting  performance. What if the volumes of reagents  pipetted  to and from the assays are only compared for precision  and not accuracy? An undetected variability  in accuracy  will impact the integrity of the assay as the automation process  continues. Even if the manual method  and automated method  both have highly precise liquid delivery,  they might not be pipetting  the same volume of reagent  to the assay.

Accuracy  information is critical to determine the deviation  of the dispensed volumes  from the target volume  when transferring or scaling-up an assay. Validating the liquid delivery steps for each assay using a robust measurement method  should be implemented to uncover  discrepancies in pipetting  performance. This presentation discusses the importance of knowing  both accuracy  and precision  information when a manual method is transferred to a Biomek®  liquid handler.  By using a standardized volume  verification methodology, it was determined that the rate-limiting reagent  was not being accurately pipetted  between  the manual and automated methods  per the protocol,  and the automation was not to blame.

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