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RNA Library Prep v1.1

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Overview

The RNA Library Prep AmpliSeq for Illumina TCR beta-SR Panel is part of the AmpliSeq for Illumina TCR beta-SR Panel protocols which include the following functionality:

  • Preconfigured AmpliSeq for Illumina TCR beta-SR Panel protocol that supports the preparation of up to 96 uniquely indexed libraries of total RNA using the AmpliSeq™ for Illumina® workflow.

  • Automated calculation of sample and buffer volumes.

  • Automated calculation or display of reagents at every step in the protocol.

  • Automatic step transition when required.Automatic placement of samples when necessary.

  • Automated assignment of QC Pass/Fail, based on user-selected threshold values.

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Protocol 1: RNA Library Prep (AmpliSeq for Illumina TCR beta-SR Panel v1.1)

Protocol Type = Sample Prep

Next Steps Configuration

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Step 1: Dilute RNA (AmpliSeq for Illumina TCR beta-SR Panel v1.1)

  • Master Step Name = Dilute RNA (AmpliSeq for Illumina v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

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Automations

chevron-rightCopy Concentration and Set Sample Volumehashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

chevron-rightCalculate Input Amount - RNAhashtag
  • Trigger Location = Record Details

  • Trigger Style = Manual button

chevron-rightCopy Desired Concentrationhashtag
  • Trigger Location = Step

  • Trigger Style = Automatic upon exit

chevron-rightSet Next Step - Advancehashtag
  • Trigger Location = Not Used

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Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

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Record Details

Group of Defaults

chevron-rightImmune Repertoire Panelhashtag
  • Directions = If enough RNA is available, dilute to intermediate concentration ~20-1000 ng/uL.

chevron-rightTCR beta-SR Panelhashtag
  • Default = If enough RNA is available, dilute to intermediate concentration ~20-1000 ng/uL.

  • Step Data

    • Group of Defaults = TCR beta-SR Panel

    • Master Step Fields

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Step 2: Qubit (AmpliSeq for Illumina TCR beta-SR Panel v1.1)

  • Master Step Name = Qubit (AmpliSeq for Illumina v1.1)

  • Step Type = Standard QC

  • Measurement Generation = Fixed, 1

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Automations

chevron-rightAssign QC flags (Qubit QC)hashtag
  • Trigger Location = Record Details

  • Trigger Style = Manual button

chevron-rightSet Next Step and Copy Concentrationhashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

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Record Details

  • Step Data (Master Step Fields)

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Step 3: Dilute RNA (AmpliSeq for Illumina TCR beta-SR Panel v1.1)

  • Master Step Name = Dilute RNA (AmpliSeq for Illumina v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

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Automations

chevron-rightCopy Concentration and Set Sample Volumehashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

chevron-rightCalculate Input Amount - RNAhashtag
  • Trigger Location = Record Details

  • Trigger Style = Manual button

chevron-rightSet Next Step - Advancehashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

chevron-rightCopy Desired Concentrationhashtag
  • Trigger Location = Step

  • Trigger Style = Automatic upon exit

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Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

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Record Details

Group of Defaults

chevron-rightImmune Repertoire Panelhashtag
  • Directions = If enough RNA is available, dilute to intermediate concentration ~20-1000 ng/uL.

chevron-rightTCR beta-SR Panelhashtag
  • Default = If enough RNA is available, dilute to intermediate concentration ~20-1000 ng/uL.

  • Step Data

    • Group of Defaults = TCR beta-SR Panel

    • Master Step Fields

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Step 4: Reverse Transcribe (AmpliSeq for Illumina TCR beta-SR Panel v1.1)

  • Master Step Name = Reverse Transcribe RNA (AmpliSeq for Illumina v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

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Automations

chevron-rightCopy Concentration, Total Volume and Input Amounthashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

chevron-rightCalculate Input Amount - TCRbeta-SRhashtag
  • Trigger Location = Record Details

  • Trigger Style = Manual button

chevron-rightSet Next Step - Advancehashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

chevron-rightCalculate Input Amount - Transcriptome Human Genome Expression Panelhashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amounthashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amount - Focus Panelhashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amount - Immune Repertoire Panelhashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amount - Immune Response Panelhashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amount - Myeloid Panelhashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amount- Custom RNA Panelhashtag
  • Trigger Location = Not Used

chevron-rightCopy Concentration, Total Volume and Input Amount - Myeloidhashtag
  • Trigger Location = Not Used

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Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

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Placement = Enabled

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

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Record Details

Group of Defaults

chevron-rightChildhood Cancer Panelhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightComprehensive Panel v3hashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightFocus Panelhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightImmune Repertoire Panelhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightImmune Response Panelhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightMyeloid Panelhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightTCR beta-SR panelhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

chevron-rightTranscriptome HGxhashtag
  • AmpliSeq ERCC RNA Spike-in Mix = No

  • Directions = If RNA was prepared from FFPE tissue and never heat-treated, heat at 80°C for 10 minutes and then cool to room temperature before proceeding to reverse transcription.

  • Step Data

    • Group of Defaults = TCR beta-SR panel

    • Master Step Fields

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Step 5: Amplify cDNA Targets (AmpliSeq for Illumina TCR beta-SR Panel v1.1)

  • Master Step Name = Amplify cDNA Targets Standard (AmpliSeq for Illumina v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

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Automations

chevron-rightSet Sample Type and Calculate Master Mix - TCR beta-SR Panelhashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

chevron-rightSet Next Step - Advancehashtag
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

chevron-rightRouting cDNA - TCRbeta SRhashtag
  • Trigger Location = Step

  • Trigger Style = Automatic upon exit

chevron-rightCopy Desired RNA Conc. & Calculate Master Mix - Custom RNA Pool 1hashtag
  • Trigger Location = Not Used

chevron-rightCopy Desired RNA Conc. & Calculate Master Mix - Custom RNA Pool 2hashtag
  • Trigger Location = Not Used

chevron-rightRouting Samples - Myeloid Panelhashtag
  • Trigger Location = Not Used

ℹ The workflow version and step version for the routing script may vary depending on the version of the IPP.

chevron-rightRouting Samples - TCRbeta SR Panelhashtag
  • Trigger Location = Not Used

ℹ The workflow version and step version for the routing script may vary depending on the version of the IPP.

chevron-rightSet Sample Type and Calculate Master Mix - Custom RNAhashtag
  • Trigger Location = Not Used

chevron-rightSet Sample Type and Calculate Master Mix - Immune Repertoire Panelhashtag
  • Trigger Location = Not Used

chevron-rightSet Sample Type and Calculate Master Mix - Immune Response Panelhashtag
  • Trigger Location = Not Used

chevron-rightSet Sample Type and Calculate Master Mix - Myeloid Panelhashtag
  • Trigger Location = Not Used

chevron-rightSet Sample Type and Calculate Master Mix - TGx Panelhashtag
  • Trigger Location = Not Used

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Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

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Record Details

  • Step Data (Master Step Fields)

Naming Convention = {InputItemName}
Sample Table (Column Headers)

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Options

Additional Options and Dropdown Items

Desired Concentration (ng/uL)

Numeric

Range = 0 To 1000

Directions

Multiline Text

Read Only

Default = If enough RNA is available, dilute to intermediate concentration ~20-50 ng/uL.

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

  • Naming Convention = {InputItemName}
    Sample Table (Column Headers)

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Container

    Container Name

    Built-in

    Container

    LIMS ID (Container)

    Built-in

    Default = Concentration

    Criteria 1 - Threshold Value

    Numeric

    Criteria 2 - Operator

    Text

    Default = <=

    Criteria 2 - Source Data Field

    Text

    Default = Concentration

    Criteria 2 - Threshold Value

    Numeric

  • Step File Placeholders

    • Log - Automatically attached

    • QC Log File - Automatically attached

    • QC Result File - Automatically attached

    • Upload File - Manually uploaded

  • Sample Table

    • Enable QC Flags = Yes

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • File Column Options

      • File Column Display = Hide

      • File Attachment Method = Auto

    • Table Columns - Global Fields

  • Naming Convention = {InputItemName}
    Sample Table (Column Headers)

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Container

    Container Name

    Built-in

    Container

    LIMS ID (Container)

    Built-in

    Options

    Additional Options and Dropdown Items

    Desired Concentration (ng/uL)

    Numeric

    Range = 0 To 1000

    Directions

    Multiline Text

    Read Only

    Default = If enough RNA is available, dilute to intermediate concentration ~20-50 ng/uL.

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

  • Naming Convention = {InputItemName}
  • Reagent Kits

    • AmpliSeq cDNA Synthesis for Illumina

      • Supplier = Illumina

      • Catalog Number = 20022654

      • Website =

  • Sample Table (Column Headers)

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Container

    Container Name

    Built-in

    Container

    LIMS ID (Container)

    Built-in

    Placement Pattern = Column
  • Destination Containers

    • 96 well plate

  • Prep Input Amount (ng) = 10

  • Recommended Prep Input (ng) = Each reaction requires 1-100 ng of DNase-treated total RNA. The recommended input is 10 ng RNA.

  • Thermal Cycler Program = RT

  • Total Samples = 0

  • Prep Input Amount (ng) = 20

  • Recommended Prep Input (ng) = Each reaction requires 1-100 ng of DNase-treated total RNA per pool. The recommended input is 10 ng RNA per pool.

  • Thermal Cycler Program = RT

  • Total Samples = 0

  • Prep Input Amount (ng) = 10

  • Recommended Prep Input (ng) = Each reaction requires 1-100 ng of DNase-treated total RNA. The recommended input is 10 ng RNA.

  • Thermal Cycler Program = RT

  • Prep Input Amount (ng) = 1000

  • Recommended Prep Input (ng) = Each reaction requires 10-1000 ng of DNase-treated total RNA. If RNA is not limiting, the recommended input is 1000 ng RNA. High RNA input amounts are recommended for rare clones.

  • Thermal Cycler Program = RT

  • Prep Input Amount (ng) = 10

  • Recommended Prep Input (ng) = Each reaction requires 1-100 ng of DNase-treated total RNA. The recommended input is 10 ng RNA.

  • Thermal Cycler Program = RT

  • Total Samples = 0

  • Prep Input Amount (ng) = 10

  • Recommended Prep Input (ng) = Each reaction requires 10-100 ng of DNase-treated total RNA. The recommended input is 10 ng RNA.

  • Thermal Cycler Program = RT

  • Total Samples = 0

  • Prep Input Amount (ng) = 1000

  • Recommended Prep Input (ng) = Each reaction requires 50-1000 ng of DNase-treated total RNA. If RNA is not limited, the recommended input is 1000 ng RNA

  • Thermal Cycler Program = RT

  • Prep Input Amount (ng) = 10

  • Recommended Prep Input (ng) = Each reaction requires 1-100 ng of DNase-treated total RNA. The recommended input is 10 ng RNA.

  • Thermal Cycler Program = RT

  • Total Samples = 0

  • Options

    Additional Options and Dropdown Items

    5X AmpliSeq cDNA Reaction Mix (uL)

    Numeric

    • Decimal Places Displayed = 1

    10X AmpliSeq RT Enzyme Mix (uL)

    Numeric

    • Decimal Places Displayed = 1

    Desired Concentration (ng/uL)

    Numeric

    Read Only

    Nuclease-free water (uL)

    Numeric

    • Decimal Places Displayed = 1

    Prep Input Amount (ng)

    Numeric

    • Default = 2

    • Range = 2 To 1000

    Prep Input Volume (uL)

    Numeric

    • Decimal Places Displayed = 2

    Recommended Prep Input (ng)

    Multiline Text

    Read Only

    • Default = Each reaction requires 1-100 ng of DNase-treated total RNA. The recommended input is 10 ng RNA.

    Thermal Cycler Program

    Text

    Read Only

    • Default = RT

    Total Samples

    Numeric

    • Default = 0

    • Decimal Places Displayed = 0

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

  • Naming Convention = {InputItemName}
  • Reagent Kits

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

      • Catalog Number = 24 - 20019101; 96 - 200191102; 384 - 200191103

      • Website =

  • ℹ The workflow version and step version for the routing script may vary depending on the version of the IPP.
    Sample Table (Column Headers)

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Container

    Container Name

    Built-in

    Container

    LIMS ID (Container)

    Built-in

    Decimal Places Displayed = 1

    Nuclease-free water (uL)

    Numeric

    Decimal Places Displayed = 2

    Thermal Cycler Program

    Text

    Default = AMP_RNA

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

  • Field Name

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Criteria 1 - Operator

    Text

    Default = >=

    Criteria 1 - Source Data Field

    Text

    Field Name

    Field Name

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    5X AmpliSeq HiFi Mix (uL)

    Numeric

    Decimal Places Displayed = 2

    5X AmpliSeq TCR beta-SR Panel (uL)

    Numeric

    Field Type

    Field Type

    Field Type

    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Concentration:: = input.::Concentration:: ; output.::Conc. Units:: = input.::Conc. Units:: ; output.::Sample Volume (uL):: = 5' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
          script:evaluateDynamicExpression \
            -t true \
            -h false \
            -exp 'output.::Total Volume (uL):: = (output.::Concentration::/step.::Desired Concentration (ng/uL)::) * output.::Sample Volume (uL):: ; \
    output.::Nuclease-free water (uL):: = output.::Total Volume (uL):: - output.::Sample Volume (uL):: ; \
    output.::Input Amount (ng):: = step.::Desired Concentration (ng/uL):: * output.::Total Volume (uL):: ' \
            -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Desired Concentration (ng/uL):: = step.::Desired Concentration (ng/uL)::' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
          script:evaluateDynamicExpression \
          -t false \
          -h false \
          -exp 'nextStep = ::ADVANCE::' \
          -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {processURI:v2} -u {username} -p {password} script:assignQC -log {compoundOutputFileLuid1} -qcResult {compoundOutputFileLuid2}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} script:evaluateDynamicExpression -t false -h false -exp 'nextStep = ::ADVANCE:: ; input.::Concentration:: = output.::Concentration:: ; input.::Conc. Units:: = output.::Conc. Units::' -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Concentration:: = input.::Concentration:: ; output.::Conc. Units:: = input.::Conc. Units:: ; output.::Sample Volume (uL):: = 5' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
          script:evaluateDynamicExpression \
            -t true \
            -h false \
            -exp 'output.::Total Volume (uL):: = (output.::Concentration::/step.::Desired Concentration (ng/uL)::) * output.::Sample Volume (uL):: ; \
    output.::Nuclease-free water (uL):: = output.::Total Volume (uL):: - output.::Sample Volume (uL):: ; \
    output.::Input Amount (ng):: = step.::Desired Concentration (ng/uL):: * output.::Total Volume (uL):: ' \
            -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
          script:evaluateDynamicExpression \
          -t false \
          -h false \
          -exp 'nextStep = ::ADVANCE::' \
          -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Desired Concentration (ng/uL):: = step.::Desired Concentration (ng/uL)::' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Concentration:: = input.::Concentration:: ; \
    output.::Conc. Units:: = input.::Conc. Units:: ; \
    output.::Total Volume (uL):: = input.::Total Volume (uL):: ; \
    output.::Input Amount (ng):: = input.::Input Amount (ng):: ; \
    step.::Desired Concentration (ng/uL):: = input.::Desired Concentration (ng/uL)::' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 7) {step.::Nuclease-free water (uL):: = (7 - step.::Prep Input Volume (uL)::) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 2 ; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
          script:evaluateDynamicExpression \
          -t false \
          -h false \
          -exp 'nextStep = ::ADVANCE::' \
          -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 3.5) {step.::Nuclease-free water (uL):: = (3.5 - (step.::Prep Input Volume (uL):: + output.::AmpliSeq ERCC RNA Spike-in Volume (uL)::)) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 1 ; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 0.5 ; \
    step.::AmpliSeq ERCC RNA Spike-in Mix (uL):: = step.::Total Samples:: * output.::AmpliSeq ERCC RNA Spike-in Volume (uL):: *1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 7) {step.::Nuclease-free water (uL):: = (7 - step.::Prep Input Volume (uL)::) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 2 ; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 7) {step.::Nuclease-free water (uL):: = (7 - step.::Prep Input Volume (uL)::) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 2; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 3.5) {step.::Nuclease-free water (uL):: = (3.5 - step.::Prep Input Volume (uL)::) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 1 ; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 0.5' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 7.0) {step.::Nuclease-free water (uL):: = (7.0 - (step.::Prep Input Volume (uL):: + output.::AmpliSeq ERCC RNA Spike-in Volume (uL)::)) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 1 ; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 0.5 ; \
    step.::AmpliSeq ERCC RNA Spike-in Mix (uL):: = step.::Total Samples:: * output.::AmpliSeq ERCC RNA Spike-in Volume (uL):: *1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 3.5) {step.::Nuclease-free water (uL):: = (3.5 - step.::Prep Input Volume (uL)::) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 1 ; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 0.5' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t true \
    -h false \
    -exp 'step.::Prep Input Volume (uL):: = step.::Prep Input Amount (ng):: / step.::Desired Concentration (ng/uL):: ; \
    if (step.::Prep Input Volume (uL):: < 7) {step.::Nuclease-free water (uL):: = (7 - step.::Prep Input Volume (uL)::) * step.::Total Samples::*1.1} else {step.::Nuclease-free water (uL):: = 0} ; \
    step.::5X AmpliSeq cDNA Reaction Mix (uL):: = step.::Total Samples:: * 1.1 * 2; \
    step.::10X AmpliSeq RT Enzyme Mix (uL):: = step.::Total Samples:: * 1.1 * 1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Concentration:: = input.::Desired Concentration (ng/uL):: ; \
    output.::Conc. Units:: = input.::Conc. Units:: ; \
    output.::Total Volume (uL):: = input.::Total Volume (uL):: ; \
    output.::Input Amount (ng):: = input.::Input Amount (ng):: ; \
    step.::Desired Concentration (ng/uL):: = input.::Desired Concentration (ng/uL)::' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Sample Type:: = ::cDNA:: ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::5X AmpliSeq TCR beta-SR Panel (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 2.0 * step.::Total Samples:: * 1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
          script:evaluateDynamicExpression \
          -t false \
          -h false \
          -exp 'nextStep = ::ADVANCE::' \
          -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -u {username} -p {password} -i {stepURI:v2} -l {compoundOutputFileLuid0} script:changeWorkflow \
    \
    --FIELD_NAME 'Sample Type' \
    --FIELD_VALUE 'cDNA' \
    --WORKFLOW 'DNA Library Prep AmpliSeq for Illumina TCR beta-SR Panel v1.1' \
    --STEP 'Transfer RNA and DNA Amplicons (AmpliSeq for Illumina TCR beta-SR Panel v1.1)' \
    --INPUTS_OR_OUTPUTS 'OUTPUTS'"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Diluted RNA Conc. (ng/uL):: = input.::Desired Final Concentration (ng/uL):: ; \
    output.::Master Mix (uL):: = 10 ; \
    step.::Total Samples:: = step.::Total Samples:: + 1 ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = step.::Total Samples:: *1.1* 4 ; \
    step.::5X AmpliSeq Custom RNA Panel (uL):: = step.::Total Samples:: * 1.1 * 4 ; \
    output.::RNA Volume (uL):: = 10 / output.::Diluted RNA Conc. (ng/uL):: ; \
    if (output.::RNA Volume (uL):: < 10) {output.::Nuclease-free water (uL):: = 20 -(output.::Master Mix (uL):: + output.::RNA Volume (uL)::)}' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Diluted RNA Conc. (ng/uL):: = input.::Desired Final Concentration (ng/uL):: ; \
    output.::Master Mix (uL):: = 10 ; \
    step.::Total Samples:: = step.::Total Samples:: + 1 ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = step.::Total Samples:: *1.1* 4.5 ; \
    step.::5X AmpliSeq Custom RNA Panel (uL):: = step.::Total Samples:: * 1.1 * 4 ; \
    output.::RNA Volume (uL):: = 10 / output.::Diluted RNA Conc. (ng/uL):: ; \
    if (output.::RNA Volume (uL):: < 10) {output.::Nuclease-free water (uL):: = 20 -(output.::Master Mix (uL):: + output.::RNA Volume (uL)::)}' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -u {username} -p {password} -i {stepURI:v2} -l {compoundOutputFileLuid0} script:changeWorkflow \
    \
    --FIELD_NAME 'Sample Type' \
    --FIELD_VALUE 'cDNA' \
    --WORKFLOW 'DNA Library Prep AmpliSeq for Illumina Myeloid Panel v1.1' \
    --STEP 'Transfer RNA and DNA Amplicons (AmpliSeq for Illumina Myeloid Panel v1.1)' \
    --INPUTS_OR_OUTPUTS 'OUTPUTS' "
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -u {username} -p {password} -i {stepURI:v2} -l {compoundOutputFileLuid0} script:changeWorkflow \
    \
    --FIELD_NAME 'Sample Type' \
    --FIELD_VALUE 'cDNA' \
    --WORKFLOW 'DNA Library Prep AmpliSeq for Illumina TCR beta-SR Panel v1.1' \
    --STEP 'Transfer RNA and DNA Amplicons (AmpliSeq for Illumina TCR beta-SR Panel v1.1)' \
    --INPUTS_OR_OUTPUTS 'OUTPUTS' "
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Sample Type:: = ::cDNA:: ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::5X AmpliSeq Custom RNA Panel (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 2.0 * step.::Total Samples:: * 1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Sample Type:: = ::cDNA:: ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::5X AmpliSeq Immune Repertoire Panel (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 7.0 * step.::Total Samples:: * 1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Sample Type:: = ::cDNA:: ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 2.0 * step.::Total Samples:: * 1.1 ; \
    step.::5x AmpliSeq Immune Response Panel (uL):: = 2.0 * step.::Total Samples:: * 1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Sample Type:: = ::cDNA:: ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 7.0 * step.::Total Samples:: * 1.1' \
    -log {compoundOutputFileLuid0}"
    bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'step.::Total Samples:: = step.::Total Samples:: + 1' \
    -log {compoundOutputFileLuid0} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} \
    script:evaluateDynamicExpression \
    -t false \
    -h false \
    -exp 'output.::Sample Type:: = ::cDNA:: ; \
    step.::5X AmpliSeq HiFi Mix (uL):: = 4.0 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 3.0 * step.::Total Samples:: * 1.1' \
    -log {compoundOutputFileLuid0}"

    Input Amount (ng)

    Numeric

    Decimal Places Displayed = 0

    Derived Sample

    Nuclease-free water (uL)

    Numeric

    Decimal Places Displayed = 2

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Sample Volume (uL)

    Numeric

    Decimal Places Displayed = 2

    Derived Sample

    Total Volume (uL)

    Numeric

    Decimal Places Displayed = 2

    Text

    Required Field

    Derived Sample

    Sample Name

    Built-in

    Measurement

    Concentration

    Numeric

    Decimal Places Displayed = 2

    Measurement

    Conc. Units

    Text

    Input Amount (ng)

    Numeric

    Decimal Places Displayed = 0

    Derived Sample

    Nuclease-free water (uL)

    Numeric

    Decimal Places Displayed = 2

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Sample Volume (uL)

    Numeric

    Decimal Places Displayed = 2

    Derived Sample

    Total Volume (uL)

    Numeric

    Decimal Places Displayed = 2

    Input Amount (ng)

    Numeric

    Decimal Places Displayed = 0

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Total Volume (uL)

    Numeric

    Decimal Places Displayed = 2

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Project

    Project Name

    Built-in

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Derived Sample

    Concentration

    Numeric

    Required Field

    Decimal Places Displayed = 2

    Derived Sample

    Conc. Units

    Text

    Required Field

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Derived Sample

    Concentration

    Numeric

    Required Field

    Decimal Places Displayed = 2

    Derived Sample

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Derived Sample

    Concentration

    Numeric

    Required Field

    Decimal Places Displayed = 2

    Derived Sample

    Conc. Units

    Text

    Required Field

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Derived Sample

    Concentration

    Numeric

    Required Field

    Decimal Places Displayed = 2

    Derived Sample

    Conc. Units

    Text

    Required Field

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

    Category

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Container

    Container Name

    Built-in

    Container

    LIMS ID (Container)

    Built-in

    www.illumina.comarrow-up-right
    www.illumina.comarrow-up-right

    Derived Sample

    Conc. Units

    Derived Sample

    Derived Sample

    Container