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SerialDilute

SerialDilute[Options]⟹UnitOperation

transfers an amount of sample into a container and dilutes it with buffers as specified.

    
A serial dilution involves taking a liquid sample and diluting it multiple times by a dilution factor into separate containers. It starts with the concentrated sample and multiple containers with the same amount of diluent. A certain amount of concentrated sample is transferred to one of these containers, and from this container, the same amount is transferred to another container. This process is repeated until the desired number of dilutions has been made. An example is performing a series of ten-fold dilutions; 1mL of concentrated sample can be transferred to a tube containing 9mL of diluent, then 1mL from this dilution would be transferred into another tube with 9mL of diluent, and so on.
    

Experimental Principles

    Figure 1.1: A specified or calculated amount is transferred from the input sample to a container with specified or calculated amounts of Diluent. After mixing, another amount is transferred from the second container to a third container, where the buffer is diluted again with a Diluent. In this example, the source sample is being diluted by factors of 10 two times in a water Diluent.
    Figure 1.2: A specified or calculated amount is transferred from the input sample to a container with specified or calculated amounts of Diluent, and optionally, a ConcentratedBuffer. After mixing, another amount is transferred from the second container to a third container with a specified amount of Diluent, and the process repeats until the desired number of dilutions is reached. In this example, the sample is being diluted by factors of 10 two times, where the Diluent is 1X PBS, and the ConcentratedBuffer is 10X PBS.
    Figure 1.3: A specified or calculated amount is transferred from the input sample to a container with specified or calculated amounts of BufferDiluent, and optionally, a ConcentratedBuffer. After mixing, another amount is transferred from the second container to a third container with a specified amount of BufferDiluent and optionally ConcentratedBuffer, and the process repeats until the desired number of dilutions is reached. The BufferDiluent here is meant to dilute the ConcentratedBuffer at each step. In this example, the sample is being diluted by factors of 10 twi times, where the BufferDiluent is water and the ConcentratedBuffer is 10X PBS.
    Figure 1.4: A chart of equations showing how the amounts used in the serial dilutions are calculated.

Instrumentation

    Super STAR

    Figure 2.1.1: An overview of the robotic workcell. It is capable of processing plate operations including incubation, shaking, and magnetic bead separation on-deck and centrifugation, filtration, and absorbance/fluorescence/luminescence measurement off-deck, with a robot arm facilitating the plate movements.
    Figure 2.1.2: Overhead view of the liquid handling instrument deck.

    Hamilton STARlet

    Figure 2.2.1: An overview of the robotic workcell. It is capable of processing plate operations including incubation, shaking, and magnetic bead separation on-deck and filtration and absorbance/fluorescence/luminescence measurement off-deck, with a robot arm facilitating the plate movements.
    Figure 2.2.2: Overhead view of the liquid handling instrument deck.

    Eppendorf Research Plus P200

    Figure 2.3.1: The diagram above lists the available pipette sizes and corresponding tip types that are available in the ECL.

    pipetus

    Figure 2.4.1: The above diagram lists the compatible serological tip sizes that are available at the ECL.

    Eppendorf Research Plus, 8-channel 1200uL

    Figure 2.5.1: 1 to 8 samples can be transferred at the same time using the 8 channel pipette.

Experiment Options

    General

    Source

    The samples that should be serially diluted.
    Default Value: Null
    Pattern Description: An object of type or subtype Object[Sample] or Object[Container] or a prepared sample.
    Programmatic Pattern: ObjectP[{Object[Sample], Object[Container]}] | _String
    Index Matches to: Source
    This is a pooled option.

    SourceLabel

    The label of the source sample that is used in the sequential dilution.
    Default Value: Automatic
    Pattern Description: A string or Null.
    Programmatic Pattern: ((Null | _String) | Automatic) | Null
    Index Matches to: Source
    This is a pooled option.

    SourceContainerLabel

    The label of the source sample's container that is used for the transfer.
    Default Value: Automatic
    Pattern Description: A string or Null.
    Programmatic Pattern: ((Null | _String) | Automatic) | Null
    Index Matches to: Source
    This is a pooled option.

    SampleOutLabel

    The label of the sample(s) that become the SamplesOut.
    Default Value: Automatic
    Pattern Description: List of one or more a string entries or Null.
    Programmatic Pattern: ((Null | {_String..}) | Automatic) | Null
    Index Matches to: Source

    ContainerOutLabel

    The label of the container that holds the sample that becomes the SamplesOut.
    Default Value: Automatic
    Pattern Description: List of one or more a string entries or Null.
    Programmatic Pattern: ((Null | {_String..}) | Automatic) | Null
    Index Matches to: Source

    DiluentLabel

    The label of the diluent added to the diluted sample.
    Default Value: Automatic
    Pattern Description: A string or Null.
    Programmatic Pattern: ((Null | _String) | Automatic) | Null
    Index Matches to: Source

    ConcentratedBufferLabel

    The label of the concentrated buffer that is diluted by the BufferDilutionFactor in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent).
    Default Value: Automatic
    Pattern Description: A string or Null.
    Programmatic Pattern: ((Null | _String) | Automatic) | Null
    Index Matches to: Source

    BufferDiluentLabel

    The label of the buffer used to dilute the sample such that ConcentratedBuffer is diluted by BufferDilutionFactor in the final solution.
    Default Value: Automatic
    Pattern Description: A string or Null.
    Programmatic Pattern: ((Null | _String) | Automatic) | Null
    Index Matches to: Source

    SerialDilutionFactors

    The factors by which you wish to reduce the concentrations starting with SamplesIn, followed by each previous dilution in the series of dilutions. This or TargetConcentrations must be provided, or an error is thrown.
    Default Value: Automatic
    Default Calculation: Can be calculated from equations in Process Diagrams.
    Pattern Description: List of one or more greater than or equal to 1 entries or Null.
    Programmatic Pattern: ((Null | {GreaterEqualP[1]..}) | Automatic) | Null
    Index Matches to: Source
    This is a pooled option.

    NumberOfSerialDilutions

    The number of times the sample is repeatedly diluted, starting with SamplesIn, followed by each previous dilution.
    Default Value: Automatic
    Default Calculation: Automatically set to the number of TargetConcentrations or SerialDilutionFactors if provided, otherwise set to 1.
    Pattern Description: Greater than or equal to 1 in increments of 1.
    Programmatic Pattern: GreaterEqualP[1, 1] | Automatic
    Index Matches to: Source

    Analyte

    The components in SamplesIn's Composition whose final concentrations (TargetConcentrations) are attained through a series of repeated dilutions.
    Default Value: Automatic
    Default Calculation: Automatically set to the first value in the Analytes field of SamplesIn, or, if not populated, to the first analyte in the Composition field of the input sample, or if none exist, the first identity model of any kind in the Composition field.
    Pattern Description: An object of type or subtype Model[Molecule], Model[Molecule, cDNA], Model[Molecule, Oligomer], Model[Molecule, Transcript], Model[Molecule, Protein], Model[Molecule, Protein, Antibody], Model[Molecule, Carbohydrate], Model[Molecule, Polymer], Model[Resin], Model[Resin, SolidPhaseSupport], Model[Lysate], Model[ProprietaryFormulation], Model[Virus], Model[Cell], Model[Cell, Mammalian], Model[Cell, Bacteria], Model[Cell, Yeast], Model[Tissue], Model[Material], or Model[Species] or Null.
    Programmatic Pattern: ((Null | ObjectP[IdentityModelTypes]) | Automatic) | Null
    Index Matches to: Source

    BufferDilutionStrategy

    BufferDilutionStrategy describes the manner in which to generate to buffer samples for each serial dilution. FromConcentrate provides ConcentratedBuffer to each well, which is then diluted with BufferDiluent to reach a final buffer concentraion of 1X, whereas Direct uses pre-diluted buffer which is already at 1X to perform the subsequent dilutions (see Equations chart in Process Diagrams).
    Default Value: Direct
    Pattern Description: Direct or FromConcentrate.
    Programmatic Pattern: Direct | FromConcentrate
    Index Matches to: Source

    TransferAmounts

    The series of volume transferred starting from SamplesIn, and going into each subsequent dilution.
    Default Value: Automatic
    Default Calculation: Can be calculated from the equations in Process Diagrams.
    Pattern Description: List of one or more greater than or equal to 0 microliters and less than or equal to 20 liters entries.
    Programmatic Pattern: {RangeP[0*Microliter, $MaxTransferVolume]..} | Automatic
    Index Matches to: Source

    Diluent

    The solution used to reduce the concentration of the SamplesIn and each subsequent dilution.
    Default Value: Automatic
    Default Calculation: Automatically set to the Solvent of SamplesIn, or Model[Sample,"milli-Q Water"] if Solvent is not defined.
    Pattern Description: An object of type or subtype Object[Sample] or Model[Sample] or a prepared sample or Null.
    Programmatic Pattern: ((Null | (ObjectP[{Object[Sample], Model[Sample]}] | _String)) | Automatic) | Null
    Index Matches to: Source

    DiluentAmount

    The amount of solution used to reduce the concentration of the SamplesIn and each subsequent dilution.
    Default Value: Automatic
    Default Calculation: Can be calculated from the equations in Process Diagrams.
    Pattern Description: List of one or more greater than or equal to 0 microliters and less than or equal to 20 liters entries.
    Programmatic Pattern: {RangeP[0*Microliter, $MaxTransferVolume]..} | Automatic
    Index Matches to: Source

    BufferDiluent

    The solution used to reduce the concentration of the ConcentratedBuffer in the first and each subsequent dilution.
    Default Value: Null
    Default Calculation: If the BufferDilutionStrategy is Direct, it will be set to Null. If not, it will automatically be set to the Solvent of ConcentratedBuffer, or Model["milli-Q Water"] if Solvent is not defined.
    Pattern Description: An object of type or subtype Object[Sample] or Model[Sample] or a prepared sample or Null.
    Programmatic Pattern: (Null | (ObjectP[{Object[Sample], Model[Sample]}] | _String)) | Null
    Index Matches to: Source

    BufferDilutionFactor

    The factor by which to reduce the concentration of the ConcentratedBuffer (but not the input sample) in all of the dilutions.
    Default Value: Automatic
    Default Calculation: Automatically set to the ConcentrationFactor of ConcentratedBuffer. If ConcentrationFactor is not defined, automatically set to 10 and throws a warning.
    Pattern Description: Greater than or equal to 1.
    Programmatic Pattern: GreaterEqualP[1] | Automatic
    Index Matches to: Source

    BufferDiluentAmount

    The amount of buffer diluent to be added to reduce the concentration of ConcentratedBuffer in each dilution.
    Default Value: Automatic
    Default Calculation: Can be calculated from the equations in Process Diagrams.
    Pattern Description: List of one or more greater than or equal to 0 microliters and less than or equal to 20 liters entries or Null.
    Programmatic Pattern: ((Null | {RangeP[0*Microliter, $MaxTransferVolume]..}) | Automatic) | Null
    Index Matches to: Source

    ConcentratedBuffer

    The buffer used in the first dilution from SamplesIn. If BufferDiluentStrategy is FromConcentrate, then it will also be used in subsequent dilutions.
    Default Value: Null
    Pattern Description: An object of type or subtype Object[Sample] or Model[Sample] or a prepared sample or Null.
    Programmatic Pattern: (Null | (ObjectP[{Object[Sample], Model[Sample]}] | _String)) | Null
    Index Matches to: Source

    ConcentratedBufferAmount

    The amount of ConcentratedBuffer used in the first dilution from SamplesIn and each subsequent dilution.
    Default Value: Automatic
    Default Calculation: Can be calculated from the equations in Additional Information.
    Pattern Description: List of one or more greater than or equal to 0 microliters and less than or equal to 20 liters entries or Null.
    Programmatic Pattern: ((Null | {RangeP[0*Microliter, 20*Liter]..}) | Automatic) | Null
    Index Matches to: Source

    DiscardFinalTransfer

    Indicates if the final wells contain the same volume as the previously diluted wells by removing TransferAmount from the final dilution.
    Default Value: False
    Pattern Description: True or False or Null.
    Programmatic Pattern: (Null | BooleanP) | Null
    Index Matches to: Source

    ContainerOut

    The desired type of container that should be used to prepare and house the diluted samples, with indices indicating grouping of samples in the same plates, if desired.
    Default Value: Automatic
    Default Calculation: Automatically set as the PreferredContainer for the AssayVolume of the sample. For plates, attempts to fill all wells of a single plate with the same model before diluting into the next.
    Pattern Description: List of one or more an object of type or subtype Model[Container] or Object[Container] or a prepared sample entries or list of one or more {Index, Container} entries.
    Programmatic Pattern: ({(ObjectP[{Model[Container], Object[Container]}] | _String)..} | {{GreaterEqualP[1, 1] | Automatic, (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}..}) | Automatic
    Index Matches to: Source

    TargetConcentrations

    Desired concentrations of Analyte in the final diluted samples after serial dilutions.
    Default Value: Automatic
    Default Calculation: In either Direct or FromConcentrate, can be calculated from the equations in Process Diagrams.
    Pattern Description: List of one or more greater than 0 molar or greater than 0 grams per liter entries or Null.
    Programmatic Pattern: ((Null | {(GreaterP[0*Molar] | GreaterP[(0*Gram)/Liter])..}) | Automatic) | Null
    Index Matches to: SerialDilutionFactors

    FinalVolume

    The final volume in each well after the series of dilutions. This must be provided, or else an error is thrown.
    Default Value: Automatic
    Default Calculation: In either Direct or FromConcentrate, can be calculated from the equations in Process Diagrams.
    Pattern Description: List of one or more greater than or equal to 0 microliters and less than or equal to 20 liters entries.
    Programmatic Pattern: {RangeP[0*Microliter, $MaxTransferVolume]..} | Automatic
    Index Matches to: SerialDilutionFactors

    DestinationWells

    The wells in which the dilutions will occur in ContainerOut if the dilutions occur in well plates.
    Default Value: Automatic
    Default Calculation: Automatically determined based on available wells and volume of liquid.
    Pattern Description: List of one or more any well from A1 to P24 entries or Null.
    Programmatic Pattern: ((Null | {WellPositionP..}) | Automatic) | Null
    Index Matches to: SerialDilutionFactors

    Preparation

    Indicates if this unit operation is carried out primarily robotically or manually. Manual unit operations are executed by a laboratory operator and robotic unit operations are executed by a liquid handling work cell.
    Default Value: Automatic
    Pattern Description: Manual or Robotic.
    Programmatic Pattern: PreparationMethodP | Automatic

    Mixing

    TransferMix

    Indicates if mixing of the samples is needed after each dilution in the series.
    Default Value: True
    Pattern Description: True or False.
    Programmatic Pattern: BooleanP
    Index Matches to: Source

    TransferMixType

    Determines which type of mixing should be performed on the diluted samples after each dispense.
    Default Value: Automatic
    Default Calculation: Automatically set based on the Volume option and size of the container in which the sample is prepared.
    Pattern Description: Pipette or Swirl or Null.
    Programmatic Pattern: ((Null | (Pipette | Swirl)) | Automatic) | Null
    Index Matches to: Source

    TransferNumberOfMixes

    Determines the number of times the sample is mixed for discrete mixing processes such as Pipette or Swirl.
    Default Value: Automatic
    Pattern Description: Greater than or equal to 0 in increments of 1 or Null.
    Programmatic Pattern: ((Null | GreaterEqualP[0, 1]) | Automatic) | Null
    Index Matches to: Source

    Incubate

    Determines if incubation of the samples should occur after the dilutions.
    Default Value: True
    Pattern Description: True or False or Null.
    Programmatic Pattern: (Null | BooleanP) | Null
    Index Matches to: Source

    IncubationTime

    Determines how long the incubation should occur for the sample after the dilutions.
    Default Value: Automatic
    Default Calculation: Automatically set to 30 minutes unless MixType is set to Pipette or Invert, in which case it is set to Null. Note that if you are mixing AND incubating and this option is specified, it must be the same as the specified value of the IncubationTime option.
    Pattern Description: Greater than or equal to 0 hours and less than or equal to 72 hours or Null or Null.
    Programmatic Pattern: ((Null | (Null | RangeP[0*Hour, 72*Hour])) | Automatic) | Null
    Index Matches to: Source

    MaxIncubationTime

    The maximum time to let the samples incubate after the dilutions.
    Default Value: Automatic
    Default Calculation: Automatically set based on the MixType if MixUntilDissolved is set to True. If MixUntilDissolved is False, resolves to Null.
    Pattern Description: Greater than or equal to 1 second and less than or equal to 72 hours or Null or Null.
    Programmatic Pattern: ((Null | (Null | RangeP[1*Second, 72*Hour])) | Automatic) | Null
    Index Matches to: Source

    Incubation

    IncubationTemperature

    The temperature at which the diluted samples will incubate after the dilutions.
    Default Value: Automatic
    Default Calculation: Automatically set to the maximum temperature allowed for the incubator and container, or Null if Incubate is set to False.
    Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 500 degrees Celsius or Null or Null.
    Programmatic Pattern: ((Null | (Null | (RangeP[$MinIncubationTemperature, $MaxIncubationTemperature] | Ambient))) | Automatic) | Null
    Index Matches to: Source

    Post Experiment

    SamplesInStorageCondition

    The non-default conditions under which the SamplesIn of this experiment should be stored after the protocol is completed. If left unset, SamplesIn will be stored according to their current StorageCondition.
    Default Value: Null
    Pattern Description: {AmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, BacteriaIncubation, MammalianIncubation, TissueCultureCellsIncubation, MicrobialCellsIncubation, MicrobialCellsShakingIncubation, YeastCellsIncubation, YeastCellsShakingIncubation, ViralIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
    Programmatic Pattern: (Null | (Alternatives[SampleStorageTypeP | Disposal])) | Null
    Index Matches to: Source

    SamplesOutStorageCondition

    The non-default conditions under which any new samples generated by this experiment should be stored after the protocol is completed. If left unset, the new samples will be stored according to their Models' DefaultStorageCondition.
    Default Value: Null
    Pattern Description: {AmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, BacteriaIncubation, MammalianIncubation, TissueCultureCellsIncubation, MicrobialCellsIncubation, MicrobialCellsShakingIncubation, YeastCellsIncubation, YeastCellsShakingIncubation, ViralIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
    Programmatic Pattern: (Null | (Alternatives[SampleStorageTypeP | Disposal])) | Null
    Index Matches to: Source

Protocol Options

    Post Experiment

    MeasureWeight

    Indicates if any solid samples that are modified in the course of the experiment should have their weights measured and updated after running the experiment. Please note that public samples are weighed regardless of the value of this option.
    Default Value: Automatic
    Pattern Description: True or False or Null.
    Programmatic Pattern: (BooleanP | Automatic) | Null

    MeasureVolume

    Indicates if any liquid samples that are modified in the course of the experiment should have their volumes measured and updated after running the experiment. Please note that public samples are volume measured regardless of the value of this option.
    Default Value: Automatic
    Pattern Description: True or False or Null.
    Programmatic Pattern: (BooleanP | Automatic) | Null

    ImageSample

    Indicates if any samples that are modified in the course of the experiment should be freshly imaged after running the experiment. Please note that public samples are imaged regardless of the value of this option.
    Default Value: Automatic
    Pattern Description: True or False or Null.
    Programmatic Pattern: (BooleanP | Automatic) | Null

Example Calls

    Basic Examples

    Generate a new manual sample preparation protocol with a labeled ContainerOut:

Preferred Input Containers

    96-well 2mL Deep Well Plate
    2mL Tube

Warnings and Errors

    Messages
Last modified on Thu 13 Jan 2022 14:51:58


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