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Comprehensive Cancer Panel

Equalizer v1.1

Overview

The Equalizer Workflow AmpliSeq for Illumina Comprehensive Cancer Panel is part of the AmpliSeq for Illumina Comprehensive Cancer Panel protocols that include the following functionality:

  • Preconfigured AmpliSeq for Illumina Comprehensive Cancer Panel protocol that supports the preparation of up to 96 uniquely indexed libraries of genomic DNA 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. A routing script that allows sequencing of libraries using any Illumina sequencing instrument.

Protocol 1: Equalizer Workflow (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

Protocol Type = Library Prep

Next Steps Configuration

Step 1: Clean Up Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Clean Up Library (AmpliSeq for Illumina v1.1)

  • Step Type = No Outputs

  • Reagent Kits

    • Agencourt AMPure XP Beads

      • Supplier = Thermo

      • Catalog Number = NC9959336; NC9933872

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    70% EtOH Prep Date

    Date

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Step 2: Amplify Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

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

  • Step Type = No Outputs

  • Reagent Kits

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

      • Website = www.illumina.com

Automations

Count Samples and Calculate Master Mix
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

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 'step.::1X Lib AMP Mix (uL):: = step.::Total Samples:: * 1.1 * 45 ; \
step.::10X Library Amp Primers (uL):: = step.::Total Samples:: * 1.1 * 5' \
      -log {compoundOutputFileLuid0}"
Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"
Calculate Total Samples and Calculate Master Mix
  • Trigger Location = Not Used

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 'step.::1X Lib Amp Mix:: = step.::Total Samples:: * 1.1 * 45 ; \ step.::10X Library Amp Primers:: = step.::Total Samples:: * 1.1 * 5' \ -log {compoundOutputFileLuid0}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Record Details

Group of Defaults

AmpliSeq for CHS V2
  • Thermal Cycler Program = AMP

Equalizer Workflow
  • Thermal Cycler Program = EQUAL

Standard Workflow
  • Thermal Cycler Program = AMP_7

  • Step Data

    • Group of Defaults = Equalizer Workflow

    • Master Step Fields

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      1X Lib AMP Mix (uL)

      Numeric

      Decimal Places Displayed = 2

      10X Library Amp Primers (uL)

      Numeric

      Decimal Places Displayed = 2

      Thermal Cycler Program

      Text Dropdown

      Custom Entries

      Presets

      • AMP_7

      • EQUAL

      • AMP

      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

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Step 3: Perform Capture and Clean Up (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Perform Capture and Clean Up (AmpliSeq for Illumina v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

  • Reagent Kits

    • AmpliSeq Library Equalizer for Illumina

      • Supplier = Illumina

      • Website = www.illumina.com

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Record Details

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Step 4: Elute Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

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

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Placement = Enabled

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

    • Placement Pattern = Column

  • Destination Containers

    • 96 well plate

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Thermal Cycler Program

    Text

    Default = ELUTE

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Standard v1.1

Overview

The Standard Workflow AmpliSeq for Illumina Comprehensive Cancer Panel is part of the AmpliSeq for Illumina Comprehensive Cancer Panel protocols that include the following functionality:

  • Preconfigured AmpliSeq for Illumina Comprehensive Cancer Panel protocol that supports the preparation of up to 96 uniquely indexed libraries of genomic DNA 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. A routing script that allows sequencing of libraries using any Illumina sequencing instrument.

Protocol 1: Standard Workflow (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

Protocol Type = Library Prep

Next Steps Configuration

Step 1: Clean Up Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Clean Up Library (AmpliSeq for Illumina v1.1)

  • Step Type = No Outputs

  • Reagent Kits

    • Agencourt AMPure XP Beads

      • Supplier = Thermo

      • Catalog Number = NC9959336; NC9933872

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    70% EtOH Prep Date

    Date

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Step 2: Amplify Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

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

  • Step Type = No Outputs

  • Reagent Kits

    • Agencourt AMPure XP Beads

      • Supplier = Thermo

      • Catalog Number = NC9959336; NC9933872

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

      • Website = www.illumina.com

Automations

Count Samples and Calculate Master Mix
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

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 'step.::1X Lib AMP Mix (uL):: = step.::Total Samples:: * 1.1 * 45 ; \
step.::10X Library Amp Primers (uL):: = step.::Total Samples:: * 1.1 * 5' \
      -log {compoundOutputFileLuid0}"
Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"
Calculate Total Samples and Calculate Master Mix
  • Trigger Location = Not Used

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 'step.::1X Lib Amp Mix:: = step.::Total Samples:: * 1.1 * 45 ; \ step.::10X Library Amp Primers:: = step.::Total Samples:: * 1.1 * 5' \ -log {compoundOutputFileLuid0}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Record Details

Group of Defaults

AmpliSeq for CHS V2
  • Thermal Cycler Program = AMP

Equalizer Workflow
  • Thermal Cycler Program = EQUAL

Standard Workflow
  • Thermal Cycler Program = AMP_7

  • Step Data

    • Group of Defaults = Standard Workflow

    • Master Step Fields

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      1X Lib AMP Mix (uL)

      Numeric

      Decimal Places Displayed = 2

      10X Library Amp Primers (uL)

      Numeric

      Decimal Places Displayed = 2

      Thermal Cycler Program

      Text Dropdown

      Custom Entries

      Presets

      • AMP_7

      • EQUAL

      • AMP

      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

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Step 3: Perform Second Clean Up (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Perform Second Clean Up (AmpliSeq for Illumina v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

  • Reagent Kits

    • Agencourt AMPure XP Beads

      • Supplier = Thermo

      • Catalog Number = NC9959336; NC9933872

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

      • Website = www.illumina.com

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • 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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    70% EtOH Prep Date

    Date

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      LIMS ID (Container)

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Project

      Project Name

      Built-in

Library Preparation v1.1

Overview

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

  • Preconfigured AmpliSeq for Illumina Comprehensive Cancer Panel protocol that supports the preparation of up to 96 uniquely indexed libraries of genomic DNA 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. A routing script that allows sequencing of libraries using any Illumina sequencing instrument.

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

Protocol Type = Library Prep

Next Steps Configuration

Step 1: Dilute DNA (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

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

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

Automations

Copy Concentration and Set Sample Volume
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

Calculate Input Amount
  • Trigger Location = Record Details

  • Trigger Style = Manual button

Set Next Step - Advance
  • Trigger Location = Not Used

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Record Details

  • Step Data (Master Step Fields)

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

Step 2: Qubit (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

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

  • Step Type = Standard QC

  • Measurement Generation = Fixed, 1

  • Naming Convention = {InputItemName}

Automations

Assign QC flags (Qubit QC)
  • Trigger Location = Record Details

  • Trigger Style = Manual button

Set Next Step and Copy Concentration
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Record Details

  • Step Data (Master Step Fields)

  • 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

Step 3: Dilute DNA to Final Concentration (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

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

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

Automations

Copy Concentration and Set Sample Volume
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

Calculate Input Amount
  • Trigger Location = Record Details

  • Trigger Style = Manual button

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Record Details

  • Step Data (Master Step Fields)

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

Step 4: Amplify Targets Replicates (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Amplify Targets Replicates (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

  • Reagent Kits

    • AmpliSeq Comprehensive Cancer Panel for Illumina

      • Supplier = Illumina

      • Catalog Number = 20019160

      • Website =

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

      • Website =

Automations

Copy Concentration & Input Amount - CCP
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon entry

Calculate Prep Input
  • Trigger Location = Record Details

  • Trigger Style = Manual button

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Record Details

Group of Defaults

CCP
  • Recommended Prep Input (ng) = This protocol supports 1-100 ng per pool. The recommended input is 10 ng high-quality DNA per pool.

  • Thermal Cycler Program = AMP_CCP

  • Step Data

    • Group of Defaults = CCP

    • Master Step Fields

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

Step 5: Partially Digested Amplicons with Pooling (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Partially Digested Amplicons with Pooling (AmpliSeq for Illumina v1.1)

  • Step Type = Pooling

  • Aliquot Generation = Fixed, 1

  • Naming Convention = {PoolName}

  • Reagent Kits

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

      • Website =

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

Route AmpliSeq RNA Samples for Plate Transfer - Comp. Panel
  • Trigger Location = Not Used

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Pooling

  • Label Uniqueness = Off

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

Placement = Enabled

  • Defaults

    • Well Sort Order = Row

    • Placement Pattern = Column

  • Destination Containers

    • 96 well plate

Record Details

  • Step Data (Master Step Fields)

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

Step 6: Ligate Indexes (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)

  • Master Step Name = Ligate Indexes (AmpliSeq for Illumina CCP v1.1)

  • Step Type = Add Labels

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}-{AppliedReagentLabels}

  • Reagent Kits

    • AmpliSeq Library PLUS for Illumina

      • Supplier = Illumina

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

      • Website =

Automations

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

Route AmpliSeq Samples - CCP
  • 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.

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Add Labels

  • Label Groups

    • AmpliSeq CD Indexes Set A for Illumina

    • AmpliSeq CD Indexes Set B for Illumina

    • AmpliSeq CD Indexes Set C for Illumina

    • AmpliSeq CD Indexes Set D for Illumina

    • AmpliSeq UD Indexes for Illumina (24 Indexes)

Record Details

  • Step Data (Master Step Fields)

  • Step File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Row

    • Table Columns - Global Fields

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.::Low TE (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}"

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Derived Sample

Waiting

Built-in

Project

Project Name

Built-in

Field Name

Field Type

Options

Additional Options and Dropdown Items

Desired Concentration (ng/uL)

Numeric

  • Range = 20 To 50

  • Decimal Places Displayed = 0

Directions

Multiline Text

Read Only

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

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Concentration

Numeric

Required Field

Decimal Places Displayed = 2

Derived Sample

Conc. Units

Numeric

Required Field

Derived Sample

Input Amount (ng)

Numeric

Decimal Places Displayed = 0

Derived Sample

Low TE (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

Project

Project Name

Built-in

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}"

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Derived Sample

Waiting

Built-in

Project

Project Name

Built-in

Field Name

Field Type

Options

Additional Options and Dropdown Items

Criteria 1 - Operator

Text

Default = >=

Criteria 1 - Source Data Field

Text

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

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Concentration

Numeric

Required Field

Decimal Places Displayed = 2

Derived Sample

Conc. Units

Text

Required Field

Derived Sample

Sample Name

Built-in

Measurement

Concentration

Numeric

Decimal Places Displayed = 2

Measurement

Conc. Units

Text

Project

Project Name

Built-in

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.::Low TE (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}"

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Derived Sample

Waiting

Built-in

Project

Project Name

Built-in

Field Name

Field Type

Options

Additional Options and Dropdown Items

Desired Concentration (ng/uL)

Numeric

  • Range = 20 To 50

  • Decimal Places Displayed = 0

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Concentration

Numeric

Required Field

Decimal Places Displayed = 2

Derived Sample

Conc. Units

Numeric

Required Field

Derived Sample

Input Amount (ng)

Numeric

Decimal Places Displayed = 0

Derived Sample

Low TE (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

Project

Project Name

Built-in

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.::Total Volume (uL):: = input.::Total Volume (uL):: ; output.::Input Amount (ng):: = input.::Input Amount (ng):: ; output.::5X AmpliSeq HiFi Mix (uL):: = 9.0'\
        -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.::Prep Input Volume (uL):: = output.::Prep Input Amount (ng):: / output.::Concentration:: ; \
if (output.::Prep Input Volume (uL):: < 13.5) {output.::Nuclease-free water (uL):: = 13.5 - output.::Prep Input 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}"

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Derived Sample

Waiting

Built-in

Project

Project Name

Built-in

Field Name

Field Type

Options

Additional Options and Dropdown Items

20x AmpliSeq Sample ID Panel For Illumina

Toggle Switch

Default = None Set

20x AmpliSeq Sample ID Panel For Illumina Lot#

Text

Instructions

Multiline Text

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.

PCR Cycles

Numeric

Decimal Places Displayed = 0

Recommended Prep Input (ng)

Multiline Text

Read Only

Default = This protocol supports 1-100 ng per pool. The recommended input is 10 ng high-quality DNA per pool.

Thermal Cycler Program

Text Dropdown

Custom Entries

Presets

  • AMP_CCP

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

5X AmpliSeq HiFi Mix (uL)

Numeric

  • Decimal Places Displayed = 1

Derived Sample

Concentration

Numeric

Required Field

  • Decimal Places Displayed = 2

Derived Sample

Conc. Units

Text

Required Field

Derived Sample

Input Amount (ng)

Numeric

  • Decimal Places Displayed = 0

Derived Sample

Nuclease-free water (uL)

Numeric

  • Decimal Places Displayed = 2

Derived Sample

Prep Input Amount (ng)

Numeric

Required Field

  • Range = 2 To 200

  • Decimal Places Displayed = 0

Derived Sample

Prep Input Volume (uL)

Numeric

  • Decimal Places Displayed = 2

Derived Sample

Sample Name

Built-in

Derived Sample

Total Volume (uL)

Numeric

  • Decimal Places Displayed = 2

Project

Project Name

Built-in

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 'Queue for Plate Transfer' \
--FIELD_VALUE 'Transfer Amplicons' \
--WORKFLOW 'DNA Library Prep AmpliSeq for Illumina Comprehensive Panel v3 v1.1' \
--STEP 'Transfer RNA and DNA Amplicons (AmpliSeq for Illumina Comprehensive Panel v3 v1.1)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' "

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Derived Sample

Waiting

Built-in

Project

Project Name

Built-in

Field Name

Field Type

Options

Additional Options and Dropdown Items

Thermal Cycler Program

Text

Default = FUPA

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Project

Project Name

Built-in

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 'AmpliSeq Workflow' \
--FIELD_VALUE 'Standard Workflow' \
--WORKFLOW 'Standard Workflow AmpliSeq for Illumina Comprehensive Cancer Panel v1.1' \
--STEP 'Clean Up Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'AmpliSeq Workflow' \
--FIELD_VALUE 'Equalizer Workflow' \
--WORKFLOW 'Equalizer Workflow AmpliSeq for Illumina Comprehensive Cancer Panel v1.1' \
--STEP 'Clean Up Library (AmpliSeq for Illumina Comprehensive Cancer Panel v1.1)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS'"

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

Sample Name

Built-in

Derived Sample

Waiting

Built-in

Project

Project Name

Built-in

Field Name

Field Type

Options

Additional Options and Dropdown Items

Thermal Cycler Program

Text

Default = LIGATE

Category

Field Name

Field Type

Options

Additional Options and Dropdown Items

Container

Container Name

Built-in

Container

LIMS ID (Container)

Built-in

Container

Well

Built-in

Derived Sample

AmpliSeq Workflow

Text Dropdown

Required Field

Preset

  • Standard Workflow

  • Equalizer Workflow

Derived Sample

Sample Name

Built-in

Project

Project Name

Built-in

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