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How do I do a serial dilution?

This article provides a step-by-step walkthrough of how to implement serial dilution on the Microlab Prep.

Requirements

If your labware is available, load it on the deck of the Prep so it can be auto-detected. If the labware is not available, you can still design the protocol by selecting labware options manually in the procedure.

  • Tips
  • Sample labware
  • Diluent labware (if adding diluent within this step)
  • Target labware (if transferring samples in this step)

Procedure

  • Version 2.0 or newer

    1. Tap New Protocol to get started. If you are adding a serial dilution to an existing protocol, skip to step 4.
    2. If your labware is available, place it on the deck and tap Detect. The Microlab Prep’s camera will scan the labware on the deck and identify it. A model of the deck layout will appear, with either the detected labware or empty sites where labware can be added.

      If the camera is not being used to detect labware, tap the Add button to manually select each piece of labware for the deck.
    3. Confirm the detected labware. Some detected labware may need to be defined with more detail, especially tips. To select the specific variant of a piece of labware, tap any site that is labeled “Select labware” and select which labware you loaded. If the correct labware is not listed, tap “Other…” to manually select from the full list of labware.

      If the camera was not used to detect labware, tap empty sites to add labware manually. When the list of labware appears, use the filters on the left side of the page to narrow down the list. If favorite labware has been added, the “Show only favorites” option will automatically be enabled. If your labware is not available, learn about requesting new labware.
    4. When the protocol overview appears, tap New Step, Advanced, then select Serial Dilution.
    5. If your Prep has 2 Independent Channels and an 8-Probe Head, select which pipetting tool to use and tap Next. Keep the number of samples, the labware, and the arrangement of the dilutions in mind when selecting a pipetting tool. For example, the 8 MPH may not be used with tubes.

      The selected pipetting tool will be used for all parts of the step, so keep any restrictions based on factors like the number of samples, the labware, and the arrangement of the dilutions in mind. For example, the 8 MPH may not be used with tubes.
    6. Enter values for the dilutions. By default, the total volume per well and the dilution factors must be defined, in addition to the total number of dilutions per sample. Tap Options to change how the dilutions are configured:
      • Dilution factors v. dilution volume: Set whether to calculate the volumes for the serial dilution by editing dilution factors directly or by editing the sample volume.
      • Discard excess of last dilution: Enable this option to dispense excess liquid from the last dilution to liquid waste.
      • Different volumes for each dilution: Allow the dilution factors (or sample volume) for each dilution to be customized.
      • Pipette diluent: Add diluent to the target wells within this step. If you disable this option, you need to either add diluent in another step before this one or add it to the target wells before the run starts.
      • Do first dilution in sample source well: Begin the serial dilution in the sample wells instead of transferring samples to the target wells before starting the dilutions.
    7. Tap Next after the dilutions have been configured.
    8. Create a diluent, or select an existing one. Skip this step if you are not pipetting diluent in the Serial Dilution step.
      1. Tap New Diluent, or select it from the diluent drop-down menu. A dialog to create the diluent appears.

        To use an existing diluent or reagent from another step, select it from the drop-down menu.
      2. Enter a name for the diluent and select its liquid type, then tap Create. If none of the liquid types are similar enough to the diluent, learn about managing unknown liquids.
      3. Select the labware and containers that will hold diluent. These containers will be treated as a single source; the channels or 8 MPH will aspirate repeatedly from one container until it is empty, then move on to the next. Tap Next when finished.
    9. Select the sample tubes/wells and tap Next.
    10. Select the target tubes/wells and tap Next. Target selection works differently for a serial dilution step. Tapping labware will fill it with as many complete sets of dilutions as possible, but the Prep will not perform more dilutions than the number of samples (and vice versa). Zooming in and selecting a container will fill in a complete set of dilutions starting with that container. If you selected the 8 MPH as the pipetting tool, your selections will be restricted to what all eight channels on the multi-probe head can pipette.

      If you selected the channels as the pipetting tool, you will have additional options to sort the dilutions by column, or to wrap the dilutions.
    11. Enter the required pipetting settings for the diluent, samples, and the dilutions. The tip type and dispense type must be set for the diluent, if you are pipetting diluent in this step. The samples and dilutions require a tip type and a liquid type to be selected.
    12. Adjust the default settings in the Tip, Aspirate, Dispense, and Liquid tabs for each settings screen (optional).
    13. Tap Finish to complete the step. Verify the protocol to check for errors. All changes are saved automatically, so you can exit the protocol without losing progress after finishing the step.

  • Version 1.0

    1. Tap New Protocol to get started. If you are adding a serial dilution to an existing protocol, skip to step 4.
    2. If your labware is available, place it on the deck and tap Detect. The Microlab Prep’s camera will scan the labware on the deck and identify it. A model of the deck layout will appear, with either the detected labware or empty sites where labware can be added.

      If the camera is not being used to detect labware, tap the Add button to manually select each piece of labware for the deck.
    3. Confirm the detected labware. Some detected labware may need to be defined with more detail, especially tips. To select the specific variant of a piece of labware, tap any sites that is labeled “Select labware” and select which labware you loaded. If the correct labware is not listed, tap “Other…” to manually select from the full list of labware.

      If the camera was not used to detect labware, tap empty sites to add labware manually. When the list of labware appears, use the filters on the left side of the page to narrow down the list. If favorite labware has been added, the “Show only favorites” option will automatically be enabled. If your labware is not available, learn about requesting new labware.
    4. When the protocol overview appears, tap New Step, Advanced, then select Serial Dilution.
    5. If your Prep has 2 Independent Channels and an 8-Probe Head, select which pipetting tool to use and tap Next. Keep the number of samples, the labware, and the arrangement of the dilutions in mind when selecting a pipetting tool. For example, the 8 MPH may not be used with tubes.
    6. Enter values for the dilutions. By default, the total volume per well and the dilution factors must be defined, in addition to the total number of dilutions per sample. Tap Options to change how the dilutions are configured:
      • Dilution factors v. dilution volume: Set whether to calculate the volumes for the serial dilution by editing dilution factors directly or by editing the sample volume.
      • Discard excess of last dilution: Enable this option to dispense excess liquid from the last dilution to liquid waste.
      • Different volumes for each dilution: Allow the dilution factors (or sample volume) for each dilution to be customized.
      • Pipette diluent: Add diluent to the target wells within this step. If you disable this option, you need to either add diluent in another step before this one or add it to the target wells before the run starts.
      • Do first dilution in sample source well: Begin the serial dilution in the sample wells instead of transferring samples to the target wells before starting the dilutions.
    7. Tap Next after the dilutions have been configured.
    8. Create a diluent, or select an existing one. Skip this step if you are not pipetting diluent in the Serial Dilution step.
      1. Tap New Diluent, or select it from the diluent drop-down menu. A dialog to create the diluent appears.

        To use an existing diluent or reagent from another step, select it from the drop-down menu.
      2. Enter a name for the diluent and select its liquid type, then tap Create. If none of the liquid types are similar enough to the diluent, learn about managing unknown liquids.
      3. Select the labware and containers that will hold diluent. These containers will be treated as a single source; the channels or 8 MPH will aspirate repeatedly from one container until it is empty, then move on to the next. Tap Next when finished.
    9. Select the sample tubes/wells and tap Next.
    10. Select the target tubes/wells and tap Next. Target selection works differently for a serial dilution step. Tapping labware will fill it with as many complete sets of dilutions as possible, but the Prep will not perform more dilutions than the number of samples (and vice versa). Zooming in and selecting a container will fill in a complete set of dilutions starting with that container. If you selected the 8 MPH as the pipetting tool, your selections will be restricted to what all eight channels on the multi-probe head can pipette.
    11. Enter the required pipetting settings for the diluent, samples, and the dilutions. The tip type and dispense type must be set for the diluent, if you are pipetting diluent in this step. The samples and dilutions require a tip type and a liquid type to be selected.
    12. Adjust the default settings in the Tip, Aspirate, Dispense, and Liquid tabs for each settings screen (optional).
    13. Tap Finish to complete the step. Verify the protocol to check for errors. All changes are saved automatically, so you can exit the protocol without losing progress after finishing the step.

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