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Equalizer v1.1

Overview

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

  • Preconfigured AmpliSeq for Illumina Immune Response 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.

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

Protocol 1: Equalizer Workflow (AmpliSeq for Illumina Immune Response Panel v1.1)

Protocol Type = Library Prep

Next Steps Configuration

Step 1: Clean Up Library (AmpliSeq for Illumina Immune Response Panel v1.1)

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

  • Step Type = No Outputs

  • Reagent Kits

    • Agencourt AMPure XP Beads

Automations

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

Step 2: Amplify Library (AmpliSeq for Illumina Immune Response Panel v1.1)

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

  • Step Type = No Outputs

Automations

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

  • Trigger Style = Automatic upon entry

Set Next Step - Advance
  • Trigger Location = Record Details

  • Trigger Style = Automatic upon exit

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

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

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

Step 3: Perform Capture and Clean Up (AmpliSeq for Illumina Immune Response 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}

Automations

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 File Placeholders

    • Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

Step 4: Elute Library (AmpliSeq for Illumina Immune Response 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

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

  • Sample Table (Column Headers)

Placement = Enabled

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Row

    • Placement Pattern = Column

Record Details

  • Step Data (Master Step Fields)

  • Step File Placeholders

Supplier = Thermo

  • Catalog Number = NC9959336; NC9933872

  • 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

    • 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

    Derived Sample

    Waiting

    Built-in

    Project

    Project Name

    Built-in

    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

  • Reagent Kits

    • AmpliSeq Library Equalizer for Illumina

      • Supplier = Illumina

      • Website = www.illumina.com

    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

    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)

  • 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

    Destination Containers

    • 96 well plate

    • 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

    70% EtOH Prep Date

    Date

    Field Name

    Field Type

    Options

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Thermal Cycler Program

    Text

    Default = ELUTE

    Additional Options and Dropdown Items

    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 '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}"
    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 '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}"
    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 '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

    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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Project

    Project Name

    Built-in

    Built-in

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    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

    Container

    Well

    Built-in

    Derived Sample

    Sample Name

    Built-in

    Project

    Project Name

    Built-in