ExperimentFPLC
ExperimentFPLC[Samples]⟹Protocol
generates a Protocol to separate Samples via fast protein liquid chromatography.
Fast protein liquid chromatography (FPLC) involves multiple aqueous mobile phases, into which, samples (usually proteins but sometimes other biomolecules) are injected and flow through the stationary phase column. Analytes pass over resin within the column and selectively adsorb based on their relative affinity, leading to differential retention of unique analyte molecules. Molecules are carried downstream for chemical property analysis and optional collection. On the whole, molecular components are separated into analyzable, collectable constituents. For instance, protein mixtures can be purified based on the size of each individual protein. FPLC, in contrast with HPLC, is characterized by much higher flow rates and aqueous mobile phases.
Experimental Principles
Figure 1.1: Procedural overview of a FPLC experiment. Step 1: During the system prime, the system is rinsed with cleaning solutions. Step 2: After the system prime, the Column is installed and equilibrated to measurement conditions. Step 3: Samples are then introduced into the flow path. Step 4: The analytes are selectively retained on the downstream Column. Step 5: Upon exit from the column, the separated analytes are analyzed according to their physical properties. Step 6: Based on the property measurement and specifications, analytes are saved in output containers, if CollectFractions ->True. Step 7: After final sample measurement, the Column is rinsed, removed from the system, and stowed. Step 8: After the column is removed, the system is rinsed with cleaning solution.
Instrumentation
AKTApurifier UPC 10
Figure 2.1.1: Buffer solutions are steadily pumped through the instrument consisting of a valve system, adsorbent column, detectors, and terminates at fraction collection or waste. Samples within the Autosampler are loaded into the Sample loop via positive displacement from the Syringe. The rotation of the valve exposes the sample loop, thereby carrying the sample downstream to the column. During an optional purge at a gradient change, the flow path between the buffer inlet and injection valve is vacated and replaced with the new buffer. Within the column, molecular constituents are separated by adsorption -- a function of the buffers, column, sample properties and temperature. Analytes are continuously carried downstream to the detectors, where they can elicit peak signals. Separated analytes can be collected based on the properties of these resulting peaks.
Figure 2.1.2: Principle of Ultraviolet absorbance detection. Filtered light (with a set wavelength) passes through the flow downstream of the column. Presence of light-absorbing molecules will result in less light transmission to the recipient Diode, thereby producing a chromatographic peak.
AKTA avant 150
Figure 2.2.1: Buffer solutions are steadily pumped through the instrument consisting of a valve system, adsorbent column, detectors, and terminates at fraction collection or waste. Samples within the Autosampler are loaded into the Sample loop via positive displacement from the Syringe. The rotation of the valve exposes the sample loop, thereby carrying the sample downstream to the column. During an optional purge at a gradient change, the flow path between the buffer inlet and injection valve is vacated and replaced with the new buffer. Within the column, molecular constituents are separated by adsorption -- a function of the buffers, column, sample properties and temperature. Analytes are continuously carried downstream to the detectors, where they can elicit peak signals. Separated analytes can be collected based on the properties of these resulting peaks.
Figure 2.2.2: Principle of Ultraviolet absorbance detection. Filtered light (with a set wavelength) passes through the flow downstream of the column. Presence of light-absorbing molecules will result in less light transmission to the recipient Diode, thereby producing a chromatographic peak.
AKTA avant 25
Figure 2.3.1: Buffer solutions are steadily pumped through the instrument consisting of a valve system, adsorbent column, detectors, and terminates at fraction collection or waste. Samples within the Autosampler are loaded into the Sample loop via positive displacement from the Syringe. The rotation of the valve exposes the sample loop, thereby carrying the sample downstream to the column. During an optional purge at a gradient change, the flow path between the buffer inlet and injection valve is vacated and replaced with the new buffer. Within the column, molecular constituents are separated by adsorption -- a function of the buffers, column, sample properties and temperature. Analytes are continuously carried downstream to the detectors, where they can elicit peak signals. Separated analytes can be collected based on the properties of these resulting peaks.
Experiment Options
General
SeparationMode
The category of method to be used to retain analytes selectively. This option is used to set the column and solvents. If multiple columns are specified, this option refers to the separation mode of the first column.
Default Calculation: Automatically set depending on the specified column(s). If no columns are specified, set to SizeExclusion.
Column
Item containing the stationary phase through which the mobile phase and input samples flow. It adsorbs and separates the molecules within the sample based on the properties of the mobile phase, Samples, and Column material. Note that multiple columns may be connected in series.
Pattern Description: An object of type or subtype Model[Item, Column] or Object[Item, Column] or list of one or more an object of type or subtype Model[Item, Column] or Object[Item, Column] entries.
Programmatic Pattern: ({ObjectP[{Model[Item, Column], Object[Item, Column]}]..} | ObjectP[{Model[Item, Column], Object[Item, Column]}]) | Automatic
MaxColumnPressure
Default Calculation: Automatically set to the total of the MaxPressure from the column model information.
Scale
Indicates if the experiment is intended purify or analyze material. Analytical indicates that specific measurements will be employed (e.g the absorbance of the flow with injected sample for a given wavelength) and is amendable for quantification.
Instrument
Default Calculation: Automatically set to Model[Instrument, FPLC, "AKTA avant 150"] if Scale -> Preparative, and Model[Instrument, FPLC, "AKTA avant 25"] if Scale -> Analytical.
Pattern Description: An object of type or subtype Model[Instrument, FPLC] or Object[Instrument, FPLC]
InjectionType
The manner of introducing sample to the flow path and Column. Autosampler entails a robotic handling device that injects directly from sample vials and/or plates. FlowInjection directly pumps the sample into the column. Superloop pumps sample into a specialized syringe/sample loop that ensures constant volume introduction onto the column.
Default Calculation: For injection volumes smaller than the instrument's sample loop, set to Autosampler. If set above 20 Milliliter or SamplePumpWash is true for any sample, set to direct flow injection; otherwise, set to Superloop.
SampleLoopVolume
The tubing that contains the sample before injection into flow path and introduction into the Column.
Default Calculation: If InjectionType is Autosampler, defaults to Instrument's autosampler's smallest sample loop that can accommodate the InjectionVolume. For InjectionType -> FlowInjection, defaults to Null, and for Superloop, set to the smallest available syringe.
Pattern Description: Greater than or equal to 50 microliters and less than or equal to 10 milliliters or Null.
Detector
The type measurement to employ. Currently, we offer Absorbance, Conductance, pH, Temperature, Pressure.
Default Calculation: Automatically set to the value in the Detectors field of the Instrument option's object.
Pattern Description: Absorbance, Pressure, Temperature, Conductance, or pH or list of one or more Absorbance, Pressure, Temperature, Conductance, or pH entries.
MixerVolume
The capacity of the chamber used to homogenize the mobile phase. Higher volume is better for more viscous mobile phase. 0 Milliliter or Null indicates to use no mixing chamber.
Default Calculation: Automatically set to the default for the Instrument (1.4 mL for the Avant 25; 5 mL for Avant 150; 0.6 mL for AKTA 10).
Pattern Description: Greater than or equal to 0 milliliters and less than or equal to 5 milliliters or Null.
FlowCell
The distance that light must travel in order to measure an absorbance signal in the liquid mobile phase. For the Avant 25 and 150 instruments, 0.5, 2, and 10 mm are available. For the AKTA 10, only 2 mm is available.
Pattern Description: Greater than or equal to 0.5 millimeters and less than or equal to 10 millimeters or Null.
BufferA
Default Calculation: Automatically set from SeparationMode or the objects specified by the Gradient option.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample.
BufferB
Default Calculation: Automatically set from SeparationMode or the objects specified by the Gradient option.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample.
BufferC
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferD
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferE
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferF
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferG
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferH
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Wavelength
The physical properties of light passed through the flow for the Absorbance Detector. Up to 3 separate wavelengths may be specified.
Default Calculation: Automatically set to 254 Nanometer if Instrument -> Model[Instrument, FPLC, "AKTApurifier UPC 10"], or {280 Nanometer, 260 Nanometer} otherwise.
Pattern Description: Greater than or equal to 190 nanometers and less than or equal to 700 nanometers in increments of 1 nanometer or list of one or more greater than or equal to 190 nanometers and less than or equal to 700 nanometers in increments of 1 nanometer entries.
Programmatic Pattern: (RangeP[190*Nanometer, 700*Nanometer, 1*Nanometer] | {RangeP[190*Nanometer, 700*Nanometer, 1*Nanometer]..}) | Automatic
NumberOfReplicates
The number of times each sample will be injected in an FPLC experiment. If Aliquot -> True, this also indicates the number of times each provided sample will be aliquoted.
Sample Parameters
SamplePumpWash
When InjectionType is FlowInjection, whether to include a pump wash step for sample pump in the prime method.
InjectionTable
The order of sample, Standard, and Blank sample loading into the Instrument during measurement and/or collection. Also includes the flushing and priming of the column.
Default Calculation: Determined to the order of input samples articulated. Standard and Blank samples are inserted based on the determination of StandardFrequency and BlankFrequency. For example, StandardFrequency -> FirstAndLast and BlankFrequency -> Null result in Standard samples injected first, then samples, and then the Standard sample set again.
Pattern Description: List of one or more {Type, Sample, InjectionType, InjectionVolume, Gradient} or {Type, Sample, InjectionType, InjectionVolume, Gradient} entries.
Programmatic Pattern: {({Standard | Sample | Blank, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Automatic, Automatic | FPLCInjectionTypeP, RangeP[1*Microliter, 4*Liter] | Automatic, ObjectP[Object[Method, Gradient]] | Automatic} | {ColumnPrime | ColumnFlush, Automatic | Null, Automatic | Null, Automatic | Null, ObjectP[Object[Method, Gradient]] | Automatic})..} | Automatic
SampleTemperature
Pattern Description: Ambient or greater than or equal to 4 degrees Celsius and less than or equal to 25 degrees Celsius.
FlowInjectionPurgeCycle
When InjectionType is FlowInjection or Superloop, whether to take the leftover sample from purging the lines in the syringe and dispensing it into the sample container. If this option is False, extra sample of up to 20 mL may be used to purge the line and discarded afterward. If this option is True, some BufferA and BufferB solutions (in the already purged line) will mix with the sample for purging and be transferred back to the sample tube.
Default Calculation: Automatically set to False when InjectionType contains FlowInjection or Superloop. Set to Null otherwise.
InjectionVolume
Default Calculation: Automatically resolves from Instrument and takes into account Instrument deadvolume and Sample volume.
SampleFlowRate
Default Calculation: Automatically set to the first FlowRate if InjectionType is FlowInjection; otherwise, set to Null.
Pattern Description: Greater than or equal to 0.001 milliliters per minute and less than or equal to 150 milliliters per minute or Null.
Programmatic Pattern: (RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)] | Automatic) | Null
FractionCollection
FractionCollectionTemperature
Default Calculation: Automatically set to the FractionCollectionTemperature of the specified FractionCollectionMethod, or Ambient otherwise.
Pattern Description: Ambient or greater than or equal to 6 degrees Celsius and less than or equal to 20 degrees Celsius or Null.
CollectFractions
Indicates whether to capture and store effluent from the Column by specific time windows (fractions).
Default Calculation: Automatically set to True if Scale is Preparative or any fraction collection options are specified.
FractionCollectionMethod
The fraction collection method object which describes the conditions for which a fraction is collected. Specific parameters of the object can be overridden by other fraction collection options.
FractionCollectionStartTime
The time at which to start column effluent capture. 0*Minute will start fraction collection with the sample application. Time in this case is the duration from the start of analyte injection.
Default Calculation: Automatically set to the second time point of the gradient domains or inherited from the FractionCollectionMethod option.
FractionCollectionEndTime
The time to end column effluent capture. Time in this case is the duration from the start of analyte injection.
Default Calculation: Automatically set to the last time point of the gradient domains or inherited from a method specified by FractionCollectionMethod option.
FractionCollectionMode
The method by which fractions collection should be triggered (peak detection, a constant threshold, or a fixed fraction duration).
Default Calculation: Automatically set to Threshold, inherited from a method specified by FractionCollectionMethod option, or implicitly from other fraction collection options.
MaxFractionVolume
Default Calculation: Automatically converted from MaxCollectionPeriod * the initial FlowRate, if that's specified. Otherwise, automatically set to 50 mL if the system can handle it, otherwise, 1.8 Milliliter or inherited from a method specified by FractionCollectionMethod option.
Pattern Description: Greater than or equal to 10 microliters and less than or equal to 250 milliliters or Null.
MaxCollectionPeriod
AbsoluteThreshold
The absorbance or conductance signal value above which fractions will always be collected. Note that if specified in absorbance, this behavior is controlled by the absorbance at the first wavelength specified with the Wavelength option.
Default Calculation: Automatically set to 500 mAU when FractionCollectionMode is Threshold or inherited from a method specified by FractionCollectionMethod option.
Programmatic Pattern: ((GreaterEqualP[0*AbsorbanceUnit] | GreaterEqualP[0*(Millisiemen/Centimeter)]) | Automatic) | Null
PeakSlope
The minimum slope rate (per second) that must be met before a peak is detected and fraction collection begins. A new peak (and new fraction) can be registered once the slope drops below this again, and collection ends when the PeakEndThreshold value is reached. Note that if specified with absorbance units, this behavior is controlled by the absorbance at the first wavelength specified with the Wavelength option.
Default Calculation: Automatically inherited from a method specified by FractionCollectionMethod option.
Programmatic Pattern: ((GreaterEqualP[0*(AbsorbanceUnit/Second)] | GreaterEqualP[0*(Millisiemen/(Centimeter*Second))]) | Automatic) | Null
PeakMinimumDuration
The least amount of time that changes in slopes or threshold must be maintained before they are registered for fraction collection. Note that this behavior is controlled by the absorbance at the first wavelength specified with the Wavelength option.
Default Calculation: Automatically set to 0 Second or inherited from a method specified by FractionCollectionMethod option.
Pattern Description: Greater than or equal to 0.02 minutes and less than or equal to 1500 minutes or Null.
PeakSlopeEnd
Default Calculation: When FractionCollectionMode is Peak, automatically set to PeakSlope or inherited from a method specified by FractionCollectionMethod option.
Programmatic Pattern: ((GreaterEqualP[0*(AbsorbanceUnit/Second)] | GreaterEqualP[0*(Millisiemen/(Centimeter*Second))]) | Automatic) | Null
PeakEndThreshold
The absorbance or conductance signal value below which the end of a peak is marked and fraction collection stops. Note that if given as an absorbance value, this behavior is controlled by the absorbance at the first wavelength specified with the Wavelength option.
Default Calculation: Automatically set to Null or inherited from a method specified by FractionCollectionMethod option.
Programmatic Pattern: ((GreaterEqualP[0*AbsorbanceUnit] | GreaterEqualP[0*(Millisiemen/Centimeter)]) | Automatic) | Null
Gradient
GradientA
The composition of BufferA within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientA->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferA in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option or implicitly resolved from the GradientB and SeparationMode options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer A Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientB
The composition of BufferB within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientB->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferB in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option or implicitly resolved from the GradientA and SeparationMode options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer B Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientC
The composition of BufferC within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientC->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferA in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option in order to have the percentages total to 100%.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer C Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientD
The composition of BufferD within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientB->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferB in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option or implicitly resolved from the GradientA and SeparationMode options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer D Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientE
The composition of BufferE within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientC->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferA in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option in order to have the percentages total to 100%.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer E Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientF
The composition of BufferF within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientB->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferB in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option or implicitly resolved from the GradientA and SeparationMode options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer F Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientG
The composition of BufferG within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientC->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferA in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option in order to have the percentages total to 100%.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer G Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
GradientH
The composition of BufferH within the flow, defined for specific time points. The composition is linearly interpolated for the intervening periods between the defined time points. For example for GradientB->{{0 Minute, 10 Percent},{30 Minute, 90 Percent}}, the percentage of BufferB in the flow will rise such that at 15 minutes, the composition should be 50*Percent.
Default Calculation: Automatically set from Gradient option or implicitly resolved from the GradientA and SeparationMode options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer H Composition} entries.
Programmatic Pattern: (RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic
Gradient
The buffer composition over time in the fluid flow. Specific parameters of an object can be overridden by specific options.
Default Calculation: Automatically set to best meet all the Gradient options (e.g. GradientA, GradientB, FlowRate, GradientStart, GradientEnd, GradientDuration, EquilibrateTime, FlushTime).
Pattern Description: An object of type or subtype Object[Method, Gradient] or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Buffer E Composition, Buffer F Composition, Buffer G Composition, Buffer H Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Flow Rate} entries.
Programmatic Pattern: (ObjectP[Object[Method, Gradient]] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic
FlowRate
The speed of the fluid through the pump. This speed is linearly interpolated such that consecutive entries of {Time, Flow Rate} will define the intervening fluid speed. For example, {{0 Minute, 0.3 Milliliter/Minute},{30 Minute, 0.5 Milliliter/Minute}} means flow rate of 0.4 Milliliter/Minute at 15 minutes into the run.
Default Calculation: Automatically set from SeparationMode and Scale or inherited from the method given in the Gradient option.
Pattern Description: Greater than or equal to 0.001 milliliters per minute and less than or equal to 150 milliliters per minute or list of one or more {Time, Flow Rate} entries.
Programmatic Pattern: (RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)] | {{GreaterEqualP[0*Minute], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic
GradientStart
A shorthand option to specify the starting Buffer B composition. This option must be specified with GradientEnd and GradientDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
GradientEnd
A shorthand option to specify the final Buffer B composition. This option must be specified with GradientStart and GradientDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
GradientDuration
SampleLoopDisconnect
For runs using the autosampler, the volume of buffer that should be flowed through the sample loop at the initial conditions to displace the sample, before the sample loop is disconnected from the flow path and the gradient begins. Null indicates that the sample loop is not disconnected from the flow path.
Default Calculation: Automatically set to Null, leaving the sample loop connected for the duration of the gradient.
Pattern Description: Greater than or equal to 0 milliliters and less than or equal to 4 liters or Null.
PreInjectionEquilibrationTime
The amount of time that buffer should be run through the system at the initial conditions before the sample is injected.
Default Calculation: Automatically set to 5 Minute if Instrument is Model[Instrument,FPLC,AKTA avant 150], otherwise set to Null.
EquilibrationTime
A shorthand option to specify the duration of equilibration at the starting buffer composition at the start of a gradient.
FlushTime
FlowDirection
The direction of the flow going through the column during the sample injection, controlled with the instrument software's plumbing settings. Forward indicates that the flow will go through the column in the direction indicated by the column manufacturer for standard operation. Reverse indicates that the flow will go through the column in the opposite direction indicated by the column manufacturer for standard operation. Reverse flow is only available on Model[Instrument, FPLC, AKTA avant 150] and Model[Instrument, FPLC, AKTA avant 25].
FullGradientChangePumpDisconnectAndPurge
When the gradients changes from 100% one buffer to 100% another buffer, if this option is set to True then, the flow path between the buffer inlet and injection valve is vacated and replaced with the new buffer.
PumpDisconnectOnFullGradientChangePurgeVolume
Default Calculation: Set to Null if FullGradientChangePumpDisconnectAndPurge is Off; otherwise, set to 15 Milliliter.
Pattern Description: Greater than or equal to 10 milliliters and less than or equal to 50 milliliters or Null.
Standard
StandardFrequency
Pattern Description: Greater than 0 in increments of 1 or None, First, Last, FirstAndLast, or GradientChange or Null.
Programmatic Pattern: (((None | First | Last | FirstAndLast | GradientChange) | GreaterP[0, 1]) | Automatic) | Null
Standard
A reference compound to inject to the instrument, often used for quantification or to check internal measurement consistency.
Default Calculation: Automatically set from SeparationMode when any other Standard option is specified, otherwise set to {}.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardInjectionType
The manner of introducing standard to the flow path and Column. Autosampler entails a robotic handling device that injects directly from sample vials and/or plates. FlowInjection directly pumps the sample into the column. Superloop pumps sample into a specialized syringe/sample loop that ensures constant volume introduction onto the column.
Default Calculation: For standard injection volumes smaller than the instrument's sample loop, set to Autosampler. If set above 20 Milliliter, set to direct flow injection; otherwise, set to Superloop.
StandardInjectionVolume
Pattern Description: Greater than or equal to 1 microliter and less than or equal to 4 liters or Null.
StandardGradient
Default Calculation: Automatically set to best meet all the StandardGradient_ options (e.g. StandardGradientA, StandardGradientB, StandardGradientDuration, StandardFlowRate).
Pattern Description: An object of type or subtype Object[Method, Gradient] or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Buffer E Composition, Buffer F Composition, Buffer G Composition, Buffer H Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Flow Rate} entries or Null.
Programmatic Pattern: ((ObjectP[Object[Method, Gradient]] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
StandardGradientA
The composition of BufferA within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from StandardGradientB options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer A Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientB
The composition of BufferB within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from StandardGradientA options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer B Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientC
The composition of BufferC within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from the other gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer C Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientD
The composition of BufferD within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from the other gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer D Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientE
The composition of BufferE within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from the other gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer E Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientF
The composition of BufferF within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from the other gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer F Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientG
The composition of BufferG within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from the other gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer G Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardGradientH
The composition of BufferH within the flow, defined for specific time points for Standard measurement.
Default Calculation: Automatically set from StandardGradient option or implicitly set from the other gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer H Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
StandardFlowRate
Default Calculation: Automatically set from SeparationMode and Scale or inherited from the method given in the StandardGradient option.
Pattern Description: Greater than or equal to 0.001 milliliters per minute and less than or equal to 150 milliliters per minute or list of one or more {Time, Flow Rate} entries or Null.
Programmatic Pattern: ((RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)] | {{GreaterEqualP[0*Minute], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
StandardGradientStart
A shorthand option to specify the starting Buffer B composition for standard samples. This option must be specified with StandardGradientEnd and StandardGradientDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
StandardGradientEnd
A shorthand option to specify the final Buffer B composition for standard samples. This option must be specified with StandardGradientStart and StandardGradientDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
StandardGradientDuration
StandardSampleLoopDisconnect
For runs using the autosampler, the volume of buffer that should be flowed through the sample loop at the initial conditions to displace the standard, before the sample loop is disconnected from the flow path and the gradient begins. Null indicates that the sample loop is not disconnected from the flow path.
Default Calculation: Automatically set to Null, leaving the sample loop connected for the duration of the gradient.
Pattern Description: Greater than or equal to 0 milliliters and less than or equal to 4 liters or Null.
StandardPreInjectionEquilibrationTime
The amount of time that buffer should be run through the system at the initial conditions before the sample is injected.
Default Calculation: Automatically set to 5 Minute if Instrument is Model[Instrument,FPLC,AKTA avant 150], otherwise set to Null.
StandardFlowDirection
The direction of the flow going through the column during the standard injection, controlled with the instrument software's plumbing settings. Forward indicates that the flow will go through the column in the direction indicated by the column manufacturer for standard operation. Reverse indicates that the flow will go through the column in the opposite direction indicated by the column manufacturer for standard operation. Reverse flow is only available on Model[Instrument, FPLC, AKTA avant 150] and Model[Instrument, FPLC, AKTA avant 25].
StandardStorageCondition
The non-default conditions under which any standards used by this experiment should be stored after the protocol is completed. If left Null, the standard samples will be stored according to their Models' DefaultStorageCondition.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
Blanks
BlankFrequency
Pattern Description: Greater than 0 in increments of 1 or None, First, Last, FirstAndLast, or GradientChange or Null.
Programmatic Pattern: (((None | First | Last | FirstAndLast | GradientChange) | GreaterP[0, 1]) | Automatic) | Null
Blank
The object(s) to inject typically as negative controls (e.g. to test effects stemming from injection, sample solvent, or buffer).
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankInjectionType
The manner of introducing blank to the flow path and Column. Autosampler entails a robotic handling device that injects directly from sample vials and/or plates. FlowInjection directly pumps the sample into the column. Superloop pumps sample into a specialized syringe/sample loop that ensures constant volume introduction onto the column.
Default Calculation: For blank injection volumes smaller than the instrument's sample loop, set to Autosampler. If set above 20 Milliliter, set to direct flow injection; otherwise, set to Superloop.
BlankInjectionVolume
Pattern Description: Greater than or equal to 1 microliter and less than or equal to 4 liters or Null.
BlankGradient
Default Calculation: Automatically set to best meet all the BlankGradient_ options (e.g. BlankGradientA, BlankGradientB, BlankGradientDuration, BlankFlowRate).
Pattern Description: An object of type or subtype Object[Method, Gradient] or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Buffer E Composition, Buffer F Composition, Buffer G Composition, Buffer H Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Flow Rate} entries or Null.
Programmatic Pattern: ((ObjectP[Object[Method, Gradient]] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
BlankGradientA
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from BlankGradientB option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer A Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientB
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from BlankGradientA option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer B Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientC
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from the Blank gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer C Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientD
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from BlankGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer D Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientE
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from the Blank gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer E Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientF
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from BlankGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer F Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientG
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from the Blank gradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer G Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankGradientH
Default Calculation: Automatically set from BlankGradient option or implicitly resolved from BlankGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer H Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
BlankFlowRate
Default Calculation: Automatically set from SelectionMode and Scale or inherited from the method given in the BlankGradient option.
Pattern Description: Greater than or equal to 0.001 milliliters per minute and less than or equal to 150 milliliters per minute or list of one or more {Time, Flow Rate} entries or Null.
Programmatic Pattern: ((RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)] | {{GreaterEqualP[0*Minute], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
BlankGradientStart
A shorthand option to specify the starting Buffer B composition for blank samples. This option must be specified with BlankGradientEnd and BlankGradientDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
BlankGradientEnd
A shorthand option to specify the final Buffer B composition for blank samples. This option must be specified with BlankGradientStart and BlankGradientDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
BlankGradientDuration
BlankSampleLoopDisconnect
For runs using the autosampler, the volume of buffer that should be flowed through the sample loop at the initial conditions to displace the blank, before the sample loop is disconnected from the flow path and the gradient begins. Null indicates that the sample loop is not disconnected from the flow path.
Default Calculation: Automatically set to Null, leaving the sample loop connected for the duration of the gradient.
Pattern Description: Greater than or equal to 0 milliliters and less than or equal to 4 liters or Null.
BlankPreInjectionEquilibrationTime
The amount of time that buffer should be run through the system at the initial conditions before the sample is injected.
Default Calculation: Automatically set to 5 Minute if Instrument is Model[Instrument,FPLC,AKTA avant 150], otherwise set to Null.
BlankFlowDirection
The direction of the flow going through the column during the blank injection, controlled with the instrument software's plumbing settings. Forward indicates that the flow will go through the column in the direction indicated by the column manufacturer for standard operation. Reverse indicates that the flow will go through the column in the opposite direction indicated by the column manufacturer for standard operation. Reverse flow is only available on Model[Instrument, FPLC, AKTA avant 150] and Model[Instrument, FPLC, AKTA avant 25].
BlankStorageCondition
The non-default conditions under which any blanks used by this experiment should be stored after the protocol is completed. If left Null, the blank samples will be stored according to their Models' DefaultStorageCondition.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
ColumnPrime
ColumnRefreshFrequency
Specify the frequency at which column flush (where solvent is flowed without injection in order to release adsorbed compounds within the column) and primes (where solvent is flowed in order to equilibrate the column) will be inserted into the order of analyte injections. An initial column prime and final column flush will be performed unless Null or None is specified.
Default Calculation: Set to Null when InjectionTable option is specified (meaning that this option is inconsequential); otherwise, set to FirstAndLast (meaning initial column prime before the measurements and final column flush after measurements.) when there is a Column. Set to None if there is no flush or prime.
Pattern Description: Greater than 0 in increments of 1 or None, FirstAndLast, First, or Last or Null.
ColumnPrimeGradient
Default Calculation: Automatically set to best meet all the ColumnPrimeGradient_ options (e.g. ColumnPrimeGradientA, ColumnPrimeGradientB, ColumnPrimeDuration, ColumnPrimeFlowRate).
Pattern Description: An object of type or subtype Object[Method, Gradient] or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Buffer E Composition, Buffer F Composition, Buffer G Composition, Buffer H Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Flow Rate} entries or Null.
Programmatic Pattern: ((ObjectP[Object[Method, Gradient]] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
ColumnPrimeGradientA
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradientB option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer A Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientB
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradientA option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer B Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientC
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer C Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientD
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer D Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientE
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer E Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientF
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer F Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientG
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer G Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeGradientH
Default Calculation: Automatically set from ColumnPrimeGradient option or implicitly resolved from ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer H Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnPrimeFlowRate
Default Calculation: Automatically set from SeparationMode and Scale or inherited from the method given in the ColumnPrimeGradient option.
Pattern Description: Greater than or equal to 0.001 milliliters per minute and less than or equal to 150 milliliters per minute or list of one or more {Time, Flow Rate} entries or Null.
Programmatic Pattern: ((RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)] | {{GreaterEqualP[0*Minute], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
ColumnPrimeStart
A shorthand option to specify the starting Buffer B composition for column prime runs. This option must be specified with ColumnPrimeEnd and ColumnPrimeDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
ColumnPrimeEnd
A shorthand option to specify the final Buffer B composition for column prime runs. This option must be specified with ColumnPrimeStart and ColumnPrimeDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
ColumnPrimeDuration
ColumnPrimePreInjectionEquilibrationTime
The amount of time that buffer should be run through the system at the initial conditions before the sample is injected.
Default Calculation: Automatically set to 5 Minute if Instrument is Model[Instrument,FPLC,AKTA avant 150], otherwise set to Null.
ColumnPrimeFlowDirection
The direction of the flow going through the column during column prime, controlled with the instrument software's plumbing settings. Forward indicates that the flow will go through the column in the direction indicated by the column manufacturer for standard operation. Reverse indicates that the flow will go through the column in the opposite direction indicated by the column manufacturer for standard operation. Reverse flow is only available on Model[Instrument, FPLC, AKTA avant 150] and Model[Instrument, FPLC, AKTA avant 25].
ColumnFlush
ColumnFlushGradient
Default Calculation: Automatically set to best meet all the ColumnFlushGradient_ options (e.g. ColumnFlushGradientA, ColumnFlushGradientB, ColumnFlushDuration, ColumnFlushFlowRate).
Pattern Description: An object of type or subtype Object[Method, Gradient] or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Buffer E Composition, Buffer F Composition, Buffer G Composition, Buffer H Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Buffer C Composition, Buffer D Composition, Flow Rate} entries or list of one or more {Time, Buffer A Composition, Buffer B Composition, Flow Rate} entries or Null.
Programmatic Pattern: ((ObjectP[Object[Method, Gradient]] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..} | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0*Percent, 100*Percent], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
ColumnFlushGradientA
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradientB option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer A Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientB
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradientA option.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer B Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientC
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer C Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientD
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer D Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientE
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer E Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientF
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer F Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientG
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer G Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushGradientH
Default Calculation: Automatically set from ColumnFlushGradient option or implicitly resolved from ColumnFlushGradient options.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or list of one or more {Time, Buffer H Composition} entries or Null.
Programmatic Pattern: ((RangeP[0*Percent, 100*Percent] | {{GreaterEqualP[0*Minute], RangeP[0*Percent, 100*Percent]}..}) | Automatic) | Null
ColumnFlushFlowRate
Default Calculation: Automatically set from SeparationMode and Scale or inherited from the method given in the ColumnFlushGradient option.
Pattern Description: Greater than or equal to 0.001 milliliters per minute and less than or equal to 150 milliliters per minute or list of one or more {Time, Flow Rate} entries or Null.
Programmatic Pattern: ((RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)] | {{GreaterEqualP[0*Minute], RangeP[0.001*(Milliliter/Minute), 150*(Milliliter/Minute)]}..}) | Automatic) | Null
ColumnFlushStart
A shorthand option to specify the starting Buffer B composition for column flush runs. This option must be specified with ColumnFlushEnd and ColumnFlushDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
ColumnFlushEnd
A shorthand option to specify the final Buffer B composition for column flush runs. This option must be specified with ColumnFlushStart and ColumnFlushDuration.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
ColumnFlushDuration
ColumnFlushPreInjectionEquilibrationTime
The amount of time that buffer should be run through the system at the initial conditions before the sample is injected.
Default Calculation: Automatically set to 5 Minute if Instrument is Model[Instrument,FPLC,AKTA avant 150], otherwise set to Null.
ColumnFlushFlowDirection
The direction of the flow going through the column during column flush, controlled with the instrument software's plumbing settings. Forward indicates that the flow will go through the column in the direction indicated by the column manufacturer for standard operation. Reverse indicates that the flow will go through the column in the opposite direction indicated by the column manufacturer for standard operation. Reverse flow is only available on Model[Instrument, FPLC, AKTA avant 150] and Model[Instrument, FPLC, AKTA avant 25].
Model Input
PreparedModelAmount
Indicates the amount of a Model[Sample] specified as input to the experiment function that will be prepared in the PreparedModelContainer. When set to All and the input model sample is not preparable, the entire amount of the input model sample that comes from one of the Products is prepared. The selected product must have both Amount and DefaultContainerModel populated, and it must not be a KitProduct. When set to All and the input model is preparable such as water, 1 Milliliter of the input model sample is prepared.
Programmatic Pattern: ((Null | (RangeP[1*Microliter, 20*Liter] | RangeP[1*Milligram, 20*Kilogram] | GreaterP[0*Unit, 1*Unit] | GreaterP[0., 1.] | All)) | Automatic) | Null
PreparedModelContainer
Indicates the container in which a Model[Sample] specified as input to the experiment function will be prepared.
Default Calculation: If PreparedModelAmount is set to All and the input model has a product associated with both Amount and DefaultContainerModel populated, automatically set to the DefaultContainerModel value in the product. Otherwise, automatically set to Model[Container, Plate, "96-well 2mL Deep Well Plate"].
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.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
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.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
Sample Prep Options
Sample Preparation
PreparatoryUnitOperations
Specifies a sequence of transferring, aliquoting, consolidating, or mixing of new or existing samples before the main experiment. These prepared samples can be used in the main experiment by referencing their defined name. For more information, please reference the documentation for ExperimentSamplePreparation.
Pattern Description: List of one or more unit Operation ManualSamplePreparation or RoboticSamplePreparation or unit Operation must match SamplePreparationP entries or Null.
Programmatic Pattern: {((ManualSamplePreparationMethodP | RoboticSamplePreparationMethodP) | SamplePreparationP)..} | Null
Preparatory Incubation
Incubate
Indicates if the SamplesIn should be incubated at a fixed temperature prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Incubation options are set. Otherwise, resolves to False.
IncubationTemperature
Temperature at which the SamplesIn should be incubated for the duration of the IncubationTime prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 500 degrees Celsius or Null.
Programmatic Pattern: ((Ambient | RangeP[$MinIncubationTemperature, $MaxIncubationTemperature]) | Automatic) | Null
IncubationTime
Duration for which SamplesIn should be incubated at the IncubationTemperature, prior to starting the experiment.
Mix
Default Calculation: Automatically resolves to True if any Mix related options are set. Otherwise, resolves to False.
MixType
Default Calculation: Automatically resolves based on the container of the sample and the Mix option.
Pattern Description: Roll, Vortex, Sonicate, Pipette, Invert, Stir, Shake, Homogenize, Swirl, Disrupt, or Nutate or Null.
MixUntilDissolved
Indicates if the mix should be continued up to the MaxIncubationTime or MaxNumberOfMixes (chosen according to the mix Type), in an attempt dissolve any solute. Any mixing/incubation will occur prior to starting the experiment.
Default Calculation: Automatically resolves to True if MaxIncubationTime or MaxNumberOfMixes is set.
MaxIncubationTime
Maximum duration of time for which the samples will be mixed while incubated in an attempt to dissolve any solute, if the MixUntilDissolved option is chosen. This occurs prior to starting the experiment.
Default Calculation: Automatically resolves based on MixType, MixUntilDissolved, and the container of the given sample.
IncubationInstrument
Default Calculation: Automatically resolves based on the options Mix, Temperature, MixType and container of the sample.
Pattern Description: An object of type or subtype Model[Instrument, Roller], Model[Instrument, OverheadStirrer], Model[Instrument, Vortex], Model[Instrument, Shaker], Model[Instrument, BottleRoller], Model[Instrument, Roller], Model[Instrument, Sonicator], Model[Instrument, HeatBlock], Model[Instrument, Homogenizer], Model[Instrument, Disruptor], Model[Instrument, Nutator], Model[Instrument, Thermocycler], Model[Instrument, EnvironmentalChamber], Model[Instrument, Pipette], Object[Instrument, Roller], Object[Instrument, OverheadStirrer], Object[Instrument, Vortex], Object[Instrument, Shaker], Object[Instrument, BottleRoller], Object[Instrument, Roller], Object[Instrument, Sonicator], Object[Instrument, HeatBlock], Object[Instrument, Homogenizer], Object[Instrument, Disruptor], Object[Instrument, Nutator], Object[Instrument, Thermocycler], Object[Instrument, EnvironmentalChamber], or Object[Instrument, Pipette] or Null.
AnnealingTime
Minimum duration for which the SamplesIn should remain in the incubator allowing the system to settle to room temperature after the IncubationTime has passed but prior to starting the experiment.
IncubateAliquotContainer
The desired type of container that should be used to prepare and house the incubation samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
IncubateAliquotDestinationWell
The desired position in the corresponding IncubateAliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
IncubateAliquot
The amount of each sample that should be transferred from the SamplesIn into the IncubateAliquotContainer when performing an aliquot before incubation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Centrifugation
Centrifuge
Indicates if the SamplesIn should be centrifuged prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Centrifuge options are set. Otherwise, resolves to False.
CentrifugeInstrument
Pattern Description: An object of type or subtype Model[Instrument, Centrifuge] or Object[Instrument, Centrifuge] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Centrifuge], Object[Instrument, Centrifuge]}] | Automatic) | Null
CentrifugeIntensity
The rotational speed or the force that will be applied to the samples by centrifugation prior to starting the experiment.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
CentrifugeTime
CentrifugeTemperature
The temperature at which the centrifuge chamber should be held while the samples are being centrifuged prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -10 degrees Celsius and less than or equal to 40 degrees Celsius or Null.
CentrifugeAliquotContainer
The desired type of container that should be used to prepare and house the centrifuge samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
CentrifugeAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
CentrifugeAliquot
The amount of each sample that should be transferred from the SamplesIn into the CentrifugeAliquotContainer when performing an aliquot before centrifugation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Filtering
Filtration
Indicates if the SamplesIn should be filter prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Filter options are set. Otherwise, resolves to False.
FiltrationType
Default Calculation: Will automatically resolve to a filtration type appropriate for the volume of sample being filtered.
FilterInstrument
Default Calculation: Will automatically resolved to an instrument appropriate for the filtration type.
Pattern Description: An object of type or subtype Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], or Object[Instrument, SyringePump] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], Object[Instrument, SyringePump]}] | Automatic) | Null
Filter
The filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Will automatically resolve to a filter appropriate for the filtration type and instrument.
Pattern Description: An object of type or subtype Model[Container, Plate, Filter], Model[Container, Vessel, Filter], or Model[Item, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Plate, Filter], Model[Container, Vessel, Filter], Model[Item, Filter]}] | Automatic) | Null
FilterMaterial
The membrane material of the filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter material for the given sample is Filtration is set to True.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
PrefilterMaterial
The material from which the prefilter filtration membrane should be made of to remove impurities from the SamplesIn prior to starting the experiment.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
FilterPoreSize
The pore size of the filter that should be used when removing impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter pore size for the given sample is Filtration is set to True.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
PrefilterPoreSize
The pore size of the filter; all particles larger than this should be removed during the filtration.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
FilterSyringe
Default Calculation: Resolves to an syringe appropriate to the volume of sample being filtered, if Filtration is set to True.
Pattern Description: An object of type or subtype Model[Container, Syringe] or Object[Container, Syringe] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container, Syringe], Object[Container, Syringe]}] | _String) | Automatic) | Null
FilterHousing
The filter housing that should be used to hold the filter membrane when filtration is performed using a standalone filter membrane.
Default Calculation: Resolve to an housing capable of holding the size of the membrane being used, if filter with Membrane FilterType is being used and Filtration is set to True.
Pattern Description: An object of type or subtype Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], or Object[Instrument, FilterBlock] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], Object[Instrument, FilterBlock]}] | Automatic) | Null
FilterIntensity
Default Calculation: Will automatically resolve to 2000 GravitationalAcceleration if FiltrationType is Centrifuge and Filtration is True.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
FilterTime
Default Calculation: Will automatically resolve to 5 Minute if FiltrationType is Centrifuge and Filtration is True.
FilterTemperature
The temperature at which the centrifuge chamber will be held while the samples are being centrifuged during filtration.
Default Calculation: Will automatically resolve to 22 Celsius if FiltrationType is Centrifuge and Filtration is True.
FilterContainerOut
The desired container filtered samples should be produced in or transferred into by the end of filtration, with indices indicating grouping of samples in the same plates, if desired.
Default Calculation: Automatically set as the PreferredContainer for the Volume of the sample. For plates, attempts to fill all wells of a single plate with the same model before using another one.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or {Index, Container} or Null.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | {GreaterEqualP[1, 1] | Automatic, (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
FilterAliquotContainer
The desired type of container that should be used to prepare and house the filter samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquot
The amount of each sample that should be transferred from the SamplesIn into the FilterAliquotContainer when performing an aliquot before filtration.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
FilterSterile
Default Calculation: Resolve to False if Filtration is indicated. If sterile filtration is desired, this option must manually be set to True.
Aliquoting
Aliquot
Indicates if aliquots should be taken from the SamplesIn and transferred into new AliquotSamples used in lieu of the SamplesIn for the experiment. Note that if NumberOfReplicates is specified this indicates that the input samples will also be aliquoted that number of times. Note that Aliquoting (if specified) occurs after any Sample Preparation (if specified).
AliquotAmount
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container if a liquid, or the current Mass or Count if a solid or counted item, respectively.
Programmatic Pattern: ((RangeP[1*Microliter, 20*Liter] | RangeP[1*Milligram, 20*Kilogram] | GreaterP[0*Unit, 1*Unit] | GreaterP[0., 1.] | All) | Automatic) | Null
TargetConcentration
The desired final concentration of analyte in the AliquotSamples after dilution of aliquots of SamplesIn with the ConcentratedBuffer and BufferDiluent which should be used in lieu of the SamplesIn for the experiment.
TargetConcentrationAnalyte
Default Calculation: Automatically set to the first value in the Analytes field of the input sample, 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.
AssayVolume
Default Calculation: Automatically determined based on Volume and TargetConcentration option values.
Pattern Description: Greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
ConcentratedBuffer
The concentrated buffer which should be diluted by the BufferDilutionFactor in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferDilutionFactor
The dilution factor by which the concentrated buffer should be diluted in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: If ConcentratedBuffer is specified, automatically set to the ConcentratedBufferDilutionFactor of that sample; otherwise, set to Null.
BufferDiluent
The buffer used to dilute the aliquot sample such that ConcentratedBuffer is diluted by BufferDilutionFactor in the final solution. The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AssayBuffer
The buffer that should be added to any aliquots requiring dilution, where the volume of this buffer added is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is not specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AliquotSampleStorageCondition
The non-default conditions under which any aliquot samples generated by this experiment should be stored after the protocol is completed.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
DestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
Pattern Description: Any well from A1 to H12 or list of one or more any well from A1 to H12 or any well from A1 to H12 entries or Null.
Programmatic Pattern: ((WellPositionP | {((Automatic | Null) | WellPositionP)..}) | Automatic) | Null
AliquotContainer
The desired type of container that should be used to prepare and house the aliquot samples, with indices indicating grouping of samples in the same plates, if desired. This option will resolve to be the length of the SamplesIn * NumberOfReplicates.
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 aliquoting into the next.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null or {Index, Container} or list of one or more an object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null entries or list of one or more Automatic or Null or {Index, Container} entries.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null) | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | {((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null))..} | {({GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | (Automatic | Null))..}) | Automatic) | Null
AliquotPreparation
Default Calculation: Automatic resolution will occur based on manipulation volumes and container types.
ConsolidateAliquots
Protocol Options
Organizational Information
Template
A template protocol whose methodology should be reproduced in running this experiment. Option values will be inherited from the template protocol, but can be individually overridden by directly specifying values for those options to this Experiment function.
Pattern Description: An object of type or subtype Object[Protocol] or an object of type or subtype of Object[Protocol] with UnresolvedOptions, ResolvedOptions specified or Null.
Programmatic Pattern: (ObjectP[Object[Protocol]] | FieldReferenceP[Object[Protocol], {UnresolvedOptions, ResolvedOptions}]) | Null
Name
A object name which should be used to refer to the output object in lieu of an automatically generated ID number.
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.
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.
ImageSample
Example Calls
Basics
Fast protein liquid chromatography (FPLC) separates primarily protein mixtures by flowing through a retentive column. To run a FPLC experiment, simply run:
Separation
Various modalities are available to elicit separation in FPLC including changing the composition of the buffers over time. For example, the precise gradient timing can be articulated:
Gradients can also be defined by specifying the change at each time point, where all of the other gradients will be filled in:
Furthermore, the manner of separation can also specified. Here, analytes are retained and eluted based off their ionic properties:
Additionally, different buffers can be specified for separate sample injections and may enable rapid scouting of ideal conditions:
Fraction Collection
ExperimentFPLC can physically collect the separated analytes, Parameters can be set to control the triggering of FractionCollection. Here fractions are collected from peaks that exceed a minimum absorbance value:
ExperimentFPLC can wholly collect everything in a given time window, can set the FractionCollectionMode to Time:
ExperimentFPLC can perform slope-based fraction collection, and limit those triggers to within a specific time window:
Standards and Blanks
ExperimentFPLC can be used to quantify analytes in a sample, in which case, a Standard sample is employed to serve as reference. A standard can be submitted before and after the injection sequence of the samples simply by running:
Likewise, a Blank sample can be used to see if there is any background from the injection process. To run a blank to occur between every 5 samples, use the following command:
Absorbance detection
Injection Table
Data Processing
Warnings and Errors
Messages (84)
BlankFrequencyNoBlanks (1)
BlankSampleLoopDisconnectOptionConflict (2)
ColumnFlushOptionInjectionTableConflict (1)
ColumnPrimeOptionInjectionTableConflict (1)
ConductivityThresholdNotSupported (1)
ConflictFractionOptionSpecification (1)
ConflictingFractionCollectionOptions (3)
ConflictingFractionOptions (1)
ConflictingPeakSlopeOptions (2)
DuplicateName (1)
FeatureUnavailableFPLCInstrument (3)
FlowRateAboveMax (1)
FPLCFlowInjectionPurgeCycle (1)
FPLCInsufficientSampleVolume (2)
FPLCTooManySamples (3)
If one of the samples/blanks/standards is injecting too much volume to use on the autosampler, the maximum number of samples is only 5:
If too many samples were specified to perform in one experiment (including blanks and standards), throw an error:
If too many samples were specified to perform in one experiment (including blanks and standards), throw an error:
FractionVolumeAboveMax (1)
GradientNotReequilibrated (1)
GradientPurgeConflict (1)
GradientShortcutAmbiguity (1)
GradientShortcutConflict (1)
GradientTooLong (1)
IncompatibleInjectionVolume (2)
InjectionTableBlankConflict (1)
InjectionTableBlankFrequencyConflict (1)
InjectionTableColumnGradientConflict (1)
InjectionTableForeignBlanks (1)
InjectionTableForeignSamples (1)
InjectionTableForeignStandards (1)
InjectionTableGradientConflict (1)
InjectionTableStandardConflict (1)
InjectionTableStandardFrequencyConflict (1)
InjectionTypeLoopConflict (1)
InjectionVolumeConflict (1)
InstrumentDoesNotContainDetector (1)
InvalidFractionCollectionEndTime (2)
InvalidFractionCollectionTemperature (1)
InvalidGradientComposition (1)
InvalidSampleTemperature (1)
LowAutosamplerFlushVolume (1)
NonBinaryFPLC (2)
ObjectDoesNotExist (4)
RecoupPurgeConflict (1)
RemovedExtraGradientEntries (1)
SampleFlowRateConflict (1)
SampleLoopDisconnectInstrumentOptionConflict (1)
SampleLoopDisconnectOptionConflict (2)
StandardFrequencyNoStandards (1)
StandardSampleLoopDisconnectOptionConflict (2)
SuperloopInjectionVolumesRounded (1)
SuperloopMixedAndMatched (1)
TooManyBuffers (1)
TooManyWavelengths (1)
Possible Issues
Last modified on Mon 18 Aug 2025 18:13:02