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RNA Library Prep v1.1

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Overview

The RNA Library Prep AmpliSeq for Illumina Childhood Cancer Panel is part of the AmpliSeq for Illumina Childhood Cancer Panel protocols that include the following functionality:

  • Preconfigured AmpliSeq for Illumina Childhood Cancer Panel protocol that supports the preparation of up to 384 uniquely indexed libraries of genomic DNA or 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.

  • There is no extraction protocol in this workflow because the samples are already extracted.

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Protocol 1: RNA Library Prep (AmpliSeq for Illumina Childhood Cancer Panel v1.1)

Protocol Type = Library Prep

Next Steps Configuration

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Step 1: Dilute RNA (AmpliSeq for Illumina Childhood Cancer 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

  • Step Data (Master Step Fields)

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Step 2: Qubit (AmpliSeq for Illumina Childhood Cancer 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 Childhood Cancer 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

  • Step Data (Master Step Fields)

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Step 4: Reverse Transcribe (AmpliSeq for Illumina Childhood Cancer 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 Amounthashtag
  • 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 - 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 - TCRbeta-SRhashtag
  • Trigger Location = Not Used

chevron-rightCalculate Input Amount - Transcriptome Human Genome Expression 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

  • Step Data (Master Step Fields)

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Step 5: Amplify cDNA Targets (AmpliSeq for Illumina Childhood Cancer Panel v1.1)

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

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 2

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Automations

chevron-rightSet Sample Type and Calculate Master Mixhashtag
  • 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 Sampleshashtag
  • Trigger Location = Step

  • Trigger Style = Automatic upon exit

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

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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

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

    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

    • 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

    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}_{OutputItemSubsetNumber}
  • Reagent Kits

    • AmpliSeq Childhood Cancer Panel for Illumina

      • Supplier = Illumina

      • Catalog Number = 20028446

      • Website =

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

  • 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 = Add 5 µl 2X AmpliSeq Comprehensive Cancer Panel Pool 1 (blue cap) to the first well, and 5 µl 2X AmpliSeq Comprehensive Cancer Panel Pool 2 (blue cap) to the second. Add 5 µl 2X AmpliSeq Comprehensive Cancer Panel Pool 3 (blue cap) to the third well, and 5 µl 2X AmpliSeq Comprehensive Cancer Panel Pool 4 (blue cap) to the fourth.

    Nuclease-free water (uL)

    Numeric

    Decimal Places Displayed = 2

    PCR Cycles

    Text

    Thermal Cycler Program

    Text

    Default = AMP_RNA

    Total Samples

    Numeric

    Default = 0

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

  • Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Desired Concentration (ng/uL)

    Numeric

    Range = 0 To 1000

    Directions

    Multiline Text

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Criteria 1 - Operator

    Text

    Default = >=

    Criteria 1 - Source Data Field

    Text

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Desired Concentration (ng/uL)

    Numeric

    Range = 0 To 1000

    Directions

    Multiline Text

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    5X AmpliSeq cDNA Reaction Mix (uL)

    Numeric

    • Decimal Places Displayed = 1

    10X AmpliSeq RT Enzyme Mix (uL)

    Numeric

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    5X AmpliSeq HiFi Mix (uL)

    Numeric

    Decimal Places Displayed = 2

    Instructions - Childhood Cancel Panel

    Multiline Text

    Read Only

    Read Only

    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):: < 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 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 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):: = 4.5 * step.::Total Samples:: * 1.1 ; \
    step.::Nuclease-free water (uL):: = 3.5 * 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 Childhood Cancer Panel v1.1' \
    --STEP 'Transfer RNA and DNA Amplicons (AmpliSeq for Illumina Childhood Cancer Panel v1.1)' \
    --INPUTS_OR_OUTPUTS 'OUTPUTS' "

    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

    Website = www.illumina.comarrow-up-right

    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