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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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 progresses
Wrong 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 work
Inconsistent mixing – Incomplete mixing at one concentration level affects all subsequent concentrations because each concentration builds upon the previous one
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 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.
Opentrons helps you automate serial dilution with open-source protocols for the OT-2 and Opentrons Flex
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