Serial Dilution

Many biological or chemical experiments involve a wide range of concentrations, and serial dilution is a common technique used in such cases. Just like direct dilution, it’s crucial to ensure accurate pipetting volumes. However, mistakes can easily occur during the multiple pipetting steps involved.

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A Common Source of Experimental Failure

Although in principle the creation of a dilution series is relatively simple, many downstream experiments fail because of errors introduced in one of three ways:

Inaccurate pipetting – Since each dilution in a series is made from the previous one, inaccurate pipettes or pipetting effects are magnified as the series progressesWrong well mistakes – Creating a serial dilution involves multiple pipetting steps to and from specific wells, following a complex pattern, so mistakes are much more common than in lab workInconsistent mixing – Incomplete mixing at one concentration level affects all subsequent concentrations because each concentration builds upon the previous one

Saving Time, Avoiding Mistakes

To avoid the tedium and errors introduced by manual pipetting, consider using a liquid handling robot. Creating a protocol is made easy using the Opentrons Protocol Library – a free collection of protocols for downloading with configuration for specific volumes. This Customizable Serial Dilution protocol for Flex results in consistent, accurate dilutions without the repetitive and labor-intensive need for manual pipetting.

Serial vs Direct Dilution

Serial dilution and direct dilution both yield a range of concentrations. However, the key distinction lies in their methods:

Serial Dilution Series: Each concentration is created by diluting the previous one. This approach allows for accurate dilutions across a broader concentration range. For instance, if a stock solution in the first column is diluted 11 times (with a factor of 3 each time), the final column would be 3¹¹ = 177,147 times more dilute.

Direct Dilution Series: All concentrations are derived by taking varying amounts from a single stock solution. These can be more straightforward to set up, and negate the risk that an error affects all subsequent concentrations.

Protocol Spotlights

Opentrons helps you automate serial dilution with open-source protocols for the OT-2 and Opentrons Flex

Customizable Serial Dilution for OT-2

Serial Dilution for Protein Quantification

Standard Serial Dilution

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