TruSeq DNA PCR-Free v2.0

Protocol 1: TruSeq DNA PCR-Free v2.0

Protocol Type = Library Prep

Next Steps Configuration

Step 1: Fragment DNA (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Fragment DNA v1.0

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

  • Reagent Kits

    • TruSeq DNA PCR-Free Sample Prep Kit - Set A, B or HT Box

      • Supplier = Illumina

      • Catalog Number = Set A: FC-121-3001, Set B: FC-121-3002, HT: FC-121-3003

Automations

Normalize gDNA (PCR Free)
  • Trigger Location = Record Details

  • Trigger Style = Manual button

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 'if ( step.::Target Insert Size (bp):: == 350 ) { output.::Sample Volume (ul):: = 1000 / output.::Concentration:: } ; if ( step.::Target Insert Size (bp):: == 550 ) { output.::Sample Volume (ul):: = 2000 / output.::Concentration:: } ; output.::RSB Volume (ul):: = 55 - output.::Sample Volume (ul):: ; output.::Target Insert Size (bp):: = step.::Target Insert Size (bp)::' -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}"
Normalize gDNA (Nano)
  • 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:http} -u {username} -p {password} script:evaluateDynamicExpression -t true -h false -exp 'if ( step.::Target Insert Size (bp):: == 350 ) { output.::Sample Volume (ul):: = 100 / output.::Concentration:: } ; if ( step.::Target Insert Size (bp):: == 550 ) { output.::Sample Volume (ul):: = 200 / output.::Concentration:: } ; output.::RSB Volume (ul):: = 52.5 - output.::Sample Volume (ul):: ; output.::Target Insert Size (bp):: = step.::Target Insert Size (bp)::' -log {compoundOutputFileLuid0}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Column

  • Sample Table

    • Column Headers

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Waiting

      Built-in

    • Expanded View Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      LIMS ID (Container)

      Built-in

      Project

      Project Name

      Built-in

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Comment

    Multiline Text

    Cycles/Burst

    Numeric Dropdown

    • Custom Entries

    • Presets

      • 200

    Duration (seconds)

    Numeric Dropdown

    • Custom Entries

    • Presets

      • 65

      • 50

      • 45

      • 25

    • Decimal Places Displayed = 0

    Duty Factor (%)

    Numeric Dropdown

    • Custom Entries

    • Presets

      • 20

      • 5

      • 10

    • Decimal Places Displayed = 0

    Intensity

    Numeric Dropdown

    • Custom Entries

    • Presets

      • 5

      • 2

    • Decimal Places Displayed = 1

    Mode

    Text Dropdown

    • Custom Entries

    • Presets

      • Frequency sweeping

      • -

    Peak/Displayed Power (W)

    Numeric Dropdown

    • Custom Entries

    • Presets

      • 50

      • 175

      • 23

      • 14

      • 9

      • 7

    • Decimal Places Displayed = 0

    Target Insert Size (bp)

    Numeric Dropdown

    • Required Field

    • Custom Entries

    • Presets

      • 350

      • 550

    • Decimal Places Displayed = 0

    Temperature (oC)

    Numeric Dropdown

    • Custom Entries

    • Presets

      • 20

      • 6

    80% EtOH Prep Date

    Date

  • Step File Placeholders

    • Log file - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Column

    • 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

      Concentration

      Numeric

      Required Field

      • Decimal Places Displayed = 2

      Derived Sample

      Conc. Units

      Text

      Required Field

      Derived Sample

      RSB Volume (ul)

      Numeric

      Read Only

      • Decimal Places Displayed = 2

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Sample Volume (uL)

      Numeric

      • Decimal Places Displayed = 2

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Project

      Project Name

      Built-in

Step 2: Repair Ends and Select Library Size (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Repair Ends v1.0

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {SubmittedSampleName}

  • Reagent Kits

    • TruSeq DNA PCR-Free Sample Prep Kit - Set A, B or HT Box

      • Supplier = Illumina

      • Catalog Number = Set A: FC-121-3001, Set B: FC-121-3002, HT: FC-121-3003

Automations

SPB Dilution
  • 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 Number of Samples:: = step.::Total Number of 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 'if(input.::Target Insert Size (bp):: == 350) { step.::SPB (ml):: = (step.::Total Number of Samples:: * 109.25) / 1000} ; if(input.::Target Insert Size (bp):: == 350) { step.::PCR Grade Water (ml):: = (step.::Total Number of Samples:: * 74.75) / 1000 } ; if(input.::Target Insert Size (bp):: == 550) { step.::SPB (ml):: = (step.::Total Number of Samples:: * 92) / 1000} ; if(input.::Target Insert Size (bp):: == 550) { step.::PCR Grade Water (ml):: = (step.::Total Number of Samples:: * 92) / 1000 } ; output.::Target Insert Size (bp):: = input.::Target Insert Size (bp)::' -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}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Column

  • Sample Table

    • Column Headers

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Derived Sample

      Waiting

      Built-in

    • Expanded View Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      LIMS ID (Container)

      Built-in

      Project

      Project Name

      Built-in

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Comment

    Multiline Text

    SPB (ml)

    Numeric

    Decimal Places Displayed = 2

    PCR Grade Water (ml)

    Numeric

    Decimal Places Displayed = 2

    Thermal Cycler Program

    Text

    Default = ERP

  • Step File Placeholders

    • Next Step Log - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Column

    • Table Columns - Global Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Project

      Project Name

      Built-in

Step 3: Adenylate 3' Ends (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Adenylate 3' Ends v2.0

  • Step Type = No Outputs

  • Reagent Kits

    • TruSeq DNA PCR-Free Sample Prep Kit - Set A, B or HT Box

      • Supplier = Illumina

      • Catalog Number = Set A: FC-121-3001, Set B: FC-121-3002, HT: FC-121-3003

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

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Derived Sample

      Waiting

      Built-in

    • Expanded View Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      LIMS ID (Container)

      Built-in

      Project

      Project Name

      Built-in

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Comment

    Multiline Text

    Thermal Cycler Program

    Text

    Default = ATAIL70

  • Step File Placeholders

    • Log File - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Column

    • 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

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Project

      Project Name

      Built-in

Step 4: Ligate Adapters (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Ligate Adapters v2.0

  • Step Type = Add Labels

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {SubmittedSampleName}

  • Reagent Kits

    • TruSeq DNA PCR-Free Sample Prep Kit - Set A, B or HT Box

      • Supplier = Illumina

      • Catalog Number = Set A: FC-121-3001, Set B: FC-121-3002, HT: FC-121-3003

Automations

Copy to Output
  • 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 'output.::Target Insert Size (bp):: = input.::Target Insert Size (bp)::' -log {compoundOutputFileLuid0}"
Set Next Step & Copy to Input
  • 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:: ; output.::Target Insert Size (bp):: = input.::Target Insert Size (bp)::'  -log {compoundOutputFileLuid0}"
Set Next Step - Advance
  • 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 'nextStep = ::ADVANCE::' \
      -log {compoundOutputFileLuid0}"

Queue/Ice Bucket

  • Defaults

    • Sample Grouping = Group by Containers

    • Well Sort Order = Column

  • Sample Table

    • Column Headers

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      Container Name

      Built-in

      Container

      Well

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Derived Sample

      Waiting

      Built-in

    • Expanded View Fields

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Container

      LIMS ID (Container)

      Built-in

      Project

      Project Name

      Built-in

Add Labels

  • Label Groups

    • TruSeq DNA HT

    • TruSeq DNA LT

Record Details

  • Step Data (Master Step Fields)

    Field Name

    Field Type

    Options

    Additional Options and Dropdown Items

    Comment

    Multiline Text

    Thermal Cycler Program

    Text

    Default = LIG

    80% EtOH Prep Date

    Date

  • Step File Placeholders

    • Log File - Automatically attached

  • Sample Table

    • Sample Display Default = Expand

    • Well Sort Order = Column

    • 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

      Reagent Name

      Built-in

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Project

      Project Name

      Built-in

Step 5: Clean Up Ligated Fragments (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Clean Up Ligated Fragments (TruSeq DNA PCR-Free v2.0.10)

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

ℹ️ The version of Clean Up Ligated Fragments master step name may be different depending on the version of IPP installed.

Automations

Set Next Step & Copy to Input
  • 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:: ; output.::Target Insert Size (bp):: = input.::Target Insert Size (bp)::'  -log {compoundOutputFileLuid0}"
Set Next Step - Advance
  • 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 '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

    Comment

    Multiline Text

    80% 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 6: Validate Libraries (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Validate Libraries (TruSeq DNA PCR-Free v2.0.10)

  • Step Type = Standard QC

  • Measurement Generation = Fixed, 4

  • Naming Convention = {SubmittedSampleName}_{OutputItemSubsetNumber} of {OutputItemSubsetTotal}

ℹ️ The version of Validate Libraries master step name may be different depending on the version of IPP installed.

Automations

Set Dilution Factor
  • 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 'if (output.name.contains(::_1 of 4::)) {output.::Dilution Factor:: = ::1:10000::} ; if (output.name.contains(::_2 of 4::)) {output.::Dilution Factor:: = ::1:10000::} ; if (output.name.contains(::_3 of 4::)) {output.::Dilution Factor:: = ::1:20000::} ; if (output.name.contains(::_4 of 4::)) {output.::Dilution Factor:: = ::1:20000::}' -log {compoundOutputFileLuid0}"
Calculate Molarity and Assign QC
  • Trigger Location = Record Details

  • Trigger Style = Manual button

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 ' if (output.::Dilution Factor:: == ::1:10000:: && input.::Target Insert Size (bp):: == 350) {output.::Molarity (nM):: = ((output.::Concentration (pM):: * 10000) * (452 / 470)) / 1000} ; if (output.::Dilution Factor:: == ::1:20000:: && input.::Target Insert Size (bp):: == 350) {output.::Molarity (nM):: = ((output.::Concentration (pM):: * 20000) * (452 / 470)) / 1000} ; if (output.::Dilution Factor:: == ::1:10000:: && input.::Target Insert Size (bp):: == 550) {output.::Molarity (nM):: = ((output.::Concentration (pM):: * 10000) * (452 / 670)) / 1000} ; if (output.::Dilution Factor:: == ::1:20000:: && input.::Target Insert Size (bp):: == 550) {output.::Molarity (nM):: = ((output.::Concentration (pM):: * 10000) * (452 / 670)) / 1000}' -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:computeReplicateAverage -src 'Molarity (nM)' -dest 'Molarity (nM)' -exclude 'Exclude' -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 true -h false -exp 'if (output.::Molarity (nM):: >= 2 && output.::Molarity (nM):: <= 75) { output.QC = true } else { output.QC = false } ; if (input.::Molarity (nM):: >= 2 && input.::Molarity (nM):: <= 75) { input.QC = true } else { input.QC = false }' -log {compoundOutputFileLuid0}"
Set Next Steps - PASS/FAIL
  • 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 true -h false -excludeControls true -exp 'if (input.QC == true) { nextStep = ::ADVANCE:: } else { nextStep = ::ESCALATE:: }' -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 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

      Category

      Field Name

      Field Type

      Options

      Additional Options and Dropdown Items

      Derived Sample

      Concentration (pM)

      Numeric

      • Decimal Places Displayed = 2

      Derived Sample

      Dilution Factor

      Text Dropdown

      • Required Field

      • Custom Entries

      • Presets

        • 1:10000

        • 1:20000

      Derived Sample

      Molarity (nM)

      Numeric

      • Decimal Places Displayed = 2

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Target Insert Size (bp)

      Numeric Dropdown

      • Custom Entries

      • Presets

        • 350

        • 550

      • Decimal Places Displayed = 0

      Measurement

      Concentration (pM)

      Numeric

      • Decimal Places Displayed = 2

      Measurement

      Dilution Factor

      Text

      • Read Only

      Measurement

      Exclude

      Toggle Switch

      • Default = None Set

      Measurement

      Molarity (nM)

      Numeric

      • Decimal Places Displayed = 2

Step 7: Normalize Libraries (TruSeq DNA PCR-Free v2.0)

  • Master Step Name = Normalize Libraries 2 v2.0.10

  • Step Type = Standard

  • Derived Sample Generation = Fixed, 1

  • Naming Convention = {InputItemName}

ℹ️ The version of Normalized Libraries 2 master step name may be different depending on the version of IPP installed.

Automations

Normalization Calculations - Option 2
  • Trigger Location = Record Details

  • Trigger Style = Manual button

bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2:http} -u {username} -p {password} script:evaluateDynamicExpression -t true -h false -exp 'output.::Molarity (nM):: = input.::Molarity (nM):: ; if (output.::Molarity (nM):: <= step.::Target Normalization (nM)::) {output.::Sample Volume (ul):: = step.::Sample Volume (ul):: ; output.::Buffer Volume (ul):: = 0 ; output.::Normalized Molarity (nM):: = output.::Molarity (nM)::} else {output.::Sample Volume (ul):: = step.::Sample Volume (ul):: ; output.::Buffer Volume (ul):: = ((output.::Molarity (nM):: * step.::Sample Volume (ul)::) / step.::Target Normalization (nM)::) - step.::Sample Volume (ul):: ; output.::Normalized Molarity (nM):: = step.::Target Normalization (nM)::}' -log {compoundOutputFileLuid0}"
Set Next Step - Remove
  • 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 = ::REMOVE::' -log {compoundOutputFileLuid0}"
Routing script - Normalize Libraries
  • Trigger Location = Step

  • Trigger Style = Automatic upon exit

bash -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 'Sequencing Instrument' \
--FIELD_VALUE 'MiSeq' \
--WORKFLOW 'MiSeq Sequencing v3.2' \
--STEP 'Library Pooling (MiSeq v3.2)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NextSeq' \
--WORKFLOW 'NextSeq 500/550 Sequencing v1.2' \
--STEP 'Library Pooling (NextSeq 500/550 v1.2)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NovaSeq 2.0' \
--WORKFLOW 'NovaSeq 6000 v2.3' \
--STEP 'Define Run Format (NovaSeq 6000 v2.3)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NovaSeq 3.0' \
--WORKFLOW 'NovaSeq 6000 v3.8' \
--STEP 'Define Run Format (NovaSeq 6000 v3.8)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NovaSeqDx' \
--WORKFLOW 'NovaSeqDx v1.2' \
--STEP 'Define Run Format (NovaSeqDx v1.2)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NextSeq 1000/2000' \
--WORKFLOW 'NextSeq 1000/2000 Sequencing v2.4' \
--STEP 'Library Pooling and Dilution (NextSeq 1000/2000 Sequencing v2.4)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NovaSeq X Series' \
--WORKFLOW 'NovaSeq X Series v1.1' \
--STEP 'Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.1)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS' \
\
--FIELD_NAME 'Sequencing Instrument' \
--FIELD_VALUE 'NextSeq 1000/2000 On-Prem' \
--WORKFLOW 'NextSeq 1000/2000 On-Prem Sequencing v1.0' \
--STEP 'Library Pooling and Dilution (NextSeq 1000/2000 On-Prem Sequencing v1.0)' \
--INPUTS_OR_OUTPUTS 'OUTPUTS'"

ℹ️ The field value and the actual version of the workflows and steps in the routing automation script may be different depending on the version of IPP installed.

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

    Comment

    Multiline Text

    Sample Volume (ul)

    Numeric

    Required Field

    • Default = 10

    • Decimal Places Displayed = 0

    Target Normalization (nM)

    Numeric

    Required Field

    • Default = 10

    • Decimal Places Displayed = 0

  • Step File Placeholders

    • Log File - Automatically attached

  • Sample Table

    • Sample Display Default = Collapse

    • 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

      Molarity (nM)

      Numeric

      Decimal Places Displayed = 2

      Derived Sample

      Buffer Volume (ul)

      Numeric

      Decimal Places Displayed = 2

      Derived Sample

      Normalized Molarity (nM)

      Numeric

      Decimal Places Displayed = 2

      Derived Sample

      Sample Name

      Built-in

      Derived Sample

      Sample Volume (ul)

      Numeric

      Decimal Places Displayed = 2

      Derived Sample

      Sequencing Instrument

      Text Dropdown

      Required Field

      Presets

      • MiSeq

      • NextSeq

      • NextSeq 1000/2000

      • NextSeq 1000/2000 On-Prem

      • NovaSeq 2.0

      • NovaSeq 3.0

      • NovaSeq X Series

      • NovaSeqDx

      Project

      Project Name

      Built-in

      ℹ️ The preset options for Derived Sample Sequencing Instrument may vary depending on the version of the IPP.

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