> For the complete documentation index, see [llms.txt](https://help.bioinsight.illumina.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://help.bioinsight.illumina.com/clarity-lims/instruments-and-integrations/miseq-i100/miseq-i100-workflow-and-validation/miseq-i100-workflow-and-validation-v2.0/miseqi100-sequencing-v2.0.md).

# MiSeq i100 Series Sequencing v2.0

## Protocol 1: MiSeq i100 Series Sequencing v2.0

Protocol Type = Sequencing

**Next Steps Configuration**

* Step(s):
  * 1: Assign Analysis Configuration Template (MiSeq i100 Series Sequencing v2.0)
    * Start Next Step = Manual
    * Assign Next Step = Automatic
  * 2: Make Bulk Pool (MiSeq i100 Series Sequencing v2.0)
    * Start Next Step = Manual
    * Assign Next Step = Automatic
  * 3: Load to Dry Cartridge (MiSeq i100 Series Sequencing v2.0)
    * Start Next Step = Manual
    * Assign Next Step = Automatic
  * 4: AUTOMATED - Sequencing Run (MiSeq i100 Series Sequencing v2.0)
    * Start Next Step = Manual
    * Assign Next Step = Automatic
  * 5: AUTOMATED - Analysis Run (MiSeq i100 Series Sequencing v2.0)
    * Assign Next Step = Manual

| **Step** | **To Step 1** | **To Step 2** | **To Step 3** | **To Step 4** | **To Step 5** |
| -------- | ------------- | ------------- | ------------- | ------------- | ------------- |
| 1        |               | ✓             | ✓             | ❌             | ❌             |
| 2        |               |               | ✓             | ❌             | ❌             |
| 3        |               |               |               | ✓             | ❌             |
| 4        |               |               |               |               | ✓             |
| 5        |               |               |               |               |               |

### Step 1: Assign Analysis Configuration Template (MiSeq i100 Series Sequencing v2.0)

* Master Step Name = Assign Analysis Configuration Template (MiSeq i100 Series Sequencing v2.0)
* Step Type = No Outputs
* Instrument Types
  * MiSeq i100 Series
    * Vendor = Illumina

{% hint style="info" %}
The Instrument Types is only applicable to the on-premise integration.
{% endhint %}

{% hint style="info" %}
The version of Assign Analysis Configuration Template (MiSeq i100 Series Sequencing) master step name may be different depending on the version of IPP installed.
{% endhint %}

#### Automations

<details>

<summary>Validate Sample Names and Register Step Started</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon entry

{% code overflow="wrap" %}

```markup
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 -exp 'submittedSamples.each { sample -> if ( !sample.name.matches( ::[A-Za-z0-9_-]+:: ) ) { fail ( ::Submitted sample :: + sample.name + :: should only have alphanumeric, dash, and underscore characters. Please rename and try again.:: ) } }' -log {compoundOutputFileLuid1} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_started.groovy'"
```

{% endcode %}

</details>

<details>

<summary>1. Retrieve ACT List</summary>

* Trigger Location = Record Details
* Trigger Style = Manual button

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/miseqi100series/automation/miseqi100series-automation.jar script:retrieve_act_list -i {stepURI:v2} -u {username} -p {password} -l {compoundOutputFileLuid1} -n 'INSTRUMENT_USED' -t true"
```

{% endcode %}

</details>

<details>

<summary>2. Retrieve ACT Information</summary>

* Trigger Location = Record Details
* Trigger Style = Manual button

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/miseqi100series/automation/miseqi100series-automation.jar script:retrieve_act_info -i {stepURI:v2} -u {username} -p {password} -l {compoundOutputFileLuid1} -a '{udf:Analysis Configuration Template}' -n 'INSTRUMENT_USED'"
```

{% endcode %}

</details>

<details>

<summary>Validate Reagent Labels and Apply Selected ACT to Samples and Set Next Step</summary>

* Trigger Location = Record Details
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
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 (!input.hasValue(::Molarity (nM)::)) { fail(::The Molarity cannot be empty.::) ; } else if (input.::Molarity (nM):: <= 0) { fail(::The Molarity cannot be negative or zero.::) ;  }' -log {compoundOutputFileLuid1} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/miseqi100series/automation/miseqi100series-automation.jar -i {stepURI:v2} -u {username} -p {password} -l {compoundOutputFileLuid1} -a '{udf:Analysis Configuration Template}' -n 'INSTRUMENT_USED' --derivedSampleInstrumentIdUdf 'Instrument ID' script:validate_indexes script:assign_act && /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 (step.::Skip Pooling:: == true) { nextStep = ::Load to Dry Cartridge (MiSeq i100 Series Sequencing v2.0):: } else { nextStep = ::Make Bulk Pool (MiSeq i100 Series Sequencing v2.0):: }' -log {compoundOutputFileLuid1}"
```

{% endcode %}

> ℹ The actual version of workflow(s) and/or step(s) used in the automation script may be different depending on the version of IPP installed.

</details>

<details>

<summary>Register Step Completed</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_completed.groovy'"
```

{% endcode %}

</details>

#### Queue

* 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       |             |                                           |

#### 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** |
  | ------------------------------- | -------------- | -------------- | ----------------------------------------- |
  | Analysis Configuration Template | Text           |                | Default = No Analysis                     |
  | Application                     | Text           | Read Only      |                                           |
  | Application Version             | Text           | Read Only      |                                           |
  | Index Adapter Kit               | Text           | Read Only      |                                           |
  | Library Prep Kit                | Text           | Read Only      |                                           |
  | Reference Genome                | Text           | Read Only      |                                           |
  | Secondary Analysis Mode         | Text           | Read Only      |                                           |
  | Skip Pooling                    | Toggle Switch  | Required Field | Default = False                           |
* 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      | Well           | Built-in       |             |                                           |
    | Derived Sample | ACT Name       | Text           | Read Only   |                                           |
    | Derived Sample | Molarity (nM)  | Numeric        |             | Decimal Places Displayed = 2              |
    | Derived Sample | Sample Name    | Built-in       |             |                                           |

### Step 2: Make Bulk Pool (MiSeq i100 Series Sequencing v2.0)

* Master Step Name = Make Bulk Pool (MiSeq i100 Series Sequencing v2.0)
* Step Type = Pooling
* Aliquot Generation = Fixed, 1
* Naming Convention = {PoolName}
* Reagent Kits
  * MiSeq i100 Series Library Denature (KLD)
  * Resuspension Buffer (RSB)

{% hint style="info" %}
The version of Make Bulk Pool (MiSeq i100 Series Sequencing) master step name may be different depending on the version of IPP installed.
{% endhint %}

#### Automations

<details>

<summary>Register Step Started</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon entry

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_started.groovy'"
```

{% endcode %}

</details>

<details>

<summary>Validate Analysis Configurations</summary>

* Trigger Location = Pooling
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
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 (input.hasValue(::Instrument ID::)) {output.::Instrument ID:: = input.::Instrument ID::}' -log {compoundOutputFileLuid1} && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/miseqi100series/automation/miseqi100series-automation.jar -i {stepURI:v2} -u {username} -p {password} -l {compoundOutputFileLuid1} -m PerPool -n 'INPUT.Instrument ID' script:validate_analysis_config script:validate_physical_logical_configurations"
```

{% endcode %}

</details>

<details>

<summary>Calculate Volumes</summary>

* Trigger Location = Record Details
* Trigger Style = Manual button

{% code overflow="wrap" %}

```markup
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} -log {compoundOutputFileLuid1} \
      script:evaluateDynamicExpression \
      -exp 'if (!input.hasValue(::Molarity (nM)::)) { return; }; if (output.hasValue(::Number of Samples in Pool::)) { output.::Number of Samples in Pool:: = output.::Number of Samples in Pool:: + 1; } else { output.::Number of Samples in Pool:: = 1; };' -t true \
      script:evaluateDynamicExpression \
      -exp 'output.::MiSeq i100 Series Flowcell Type:: = step.::Flowcell Type::; \
      input.::Per Sample Volume (ul):: = 0.01 * step.::Final Loading Concentration (pM):: * step.::Bulk Pool Volume (ul):: / input.::Molarity (nM):: / output.::Number of Samples in Pool::; \
      output.::Bulk Pool Volume (ul):: = step.::Bulk Pool Volume (ul)::; \
      if (step.::% PhiX Spike In:: == 0) { step.::RSB Volume for PhiX Stock (ul):: = 0; output.::Diluted PhiX Volume (ul):: = 0; }; \
      else if (step.::% PhiX Spike In:: < 10) { step.::RSB Volume for PhiX Stock (ul):: = (10 / step.::Final Loading Concentration (pM):: * 1000 / 6) - 1; output.::Diluted PhiX Volume (ul):: = 30 * step.::% PhiX Spike In:: / (60 + 0.4 * step.::% PhiX Spike In::) }; \
      else { step.::RSB Volume for PhiX Stock (ul):: = (20 / step.::Final Loading Concentration (pM):: * 1000 / 10) - 2; output.::Diluted PhiX Volume (ul):: = 3 * step.::% PhiX Spike In:: / 10 };' -t true \
      script:calculate_multipool_adjusted_per_sample_volume -t \
      script:evaluateDynamicExpression \
      -exp 'if (output.hasValue(::Total Sample Volume (ul)::)) { output.::Total Sample Volume (ul):: = output.::Total Sample Volume (ul):: + input.::Adjusted Per Sample Volume (ul)::; } else { output.::Total Sample Volume (ul):: = input.::Adjusted Per Sample Volume (ul)::; };' -t true \
      script:evaluateDynamicExpression \
      -exp 'if (output.::Bulk Pool Volume (ul):: > output.::Total Sample Volume (ul)::) { output.::RSB Volume (ul):: = output.::Bulk Pool Volume (ul):: - output.::Total Sample Volume (ul):: } else { output.::RSB Volume (ul):: = 0 }; \
      output.::Diluted Bulk Pool Volume (ul):: = 30 - output.::Diluted PhiX Volume (ul):: ; \
      output.::KLD Volume (ul):: = 270; \
      output.::Final Loading Concentration (pM):: = step.::Final Loading Concentration (pM)::;
      output.::Final Loading Volume (ul):: = step.::Final Loading Volume (ul)::; ' -t true \
      && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/DriverFileGenerator.jar -i {stepURI:v2} -u {username} -p {password} -l {compoundOutputFileLuid1} script:driver_file_generator \
      -t /opt/gls/clarity/extensions/conf/driverfiletemplates/MiSeqi100Series_Bulk_Pool1.csv -o 1.csv \
      script:driver_file_generator \
      -t /opt/gls/clarity/extensions/conf/driverfiletemplates/MiSeqi100Series_Bulk_Pool2.csv -o 2.csv \
      script:driver_file_generator \
      -t /opt/gls/clarity/extensions/conf/driverfiletemplates/MiSeqi100Series_Bulk_Pool3.csv -o 3.csv \
      script:driver_file_generator \
      -t /opt/gls/clarity/extensions/conf/driverfiletemplates/MiSeqi100Series_Bulk_Pool4.csv -o 4.csv \
      && if [ {udf:% PhiX Spike In} != '0' ]; then \
      cat 1.csv 2.csv 3.csv 4.csv > {compoundOutputFileLuid0}.csv; \
      else \
      cat 1.csv 2.csv 4.csv > {compoundOutputFileLuid0}.csv; \
      fi \
      && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar -i {stepURI:v2} -u {username} -p {password} -log {compoundOutputFileLuid1} \
      script:evaluateDynamicExpression \
      -exp 'output.::Number of Samples in Pool:: = ::::; output.::Total Sample Volume (ul):: = ::::;' -t true \
      && echo 'Calculate Volume calculated successfully.'"
```

{% endcode %}

</details>

<details>

<summary>Set Next Step</summary>

* Trigger Location = Record Details
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
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} -log {compoundOutputFileLuid1} \
      script:evaluateDynamicExpression -exp 'if (!input.hasValue(::Molarity (nM)::)){ return; }; nextStep = ::ADVANCE::;'"
```

{% endcode %}

</details>

<details>

<summary>Register Pools and Register Step Completed</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar -i {stepURI:v2} -u {username} -p {password} script:executeUPAAutomationScript -s 'com/illumina/upa/scripts/common/pooling.groovy' script:executeUPAAutomationScript -s 'com/illumina/upa/scripts/common/step_completed.groovy'"
```

{% endcode %}

</details>

#### Queue

* 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       |             |                                           |

#### 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       |             |                                           |

#### Pooling

* Label Uniqueness = On
* Defaults
  * Sample Grouping = Group by Containers
  * Well Sort Order = Row

#### Record Details

* Step Data (Master Step Fields)

  | **Field Name**                   | **Field Type** | **Options**                                        | **Additional Options and Dropdown Items**                               |
  | -------------------------------- | -------------- | -------------------------------------------------- | ----------------------------------------------------------------------- |
  | % PhiX Spike In                  | Numeric        | Required Field                                     | <ul><li>Default = 0</li><li>Decimal Places Displayed = 1</li></ul>      |
  | Bulk Pool Volume (ul)            | Numeric        | Required Field                                     | <ul><li>Default = 30</li><li>Decimal Places Displayed = 1</li></ul>     |
  | Final Loading Concentration (pM) | Numeric        | Required Field                                     | Decimal Places Displayed = 0                                            |
  | Final Loading Volume (ul)        | Numeric        | <ul><li>Required Field</li><li>Read Only</li></ul> | <ul><li>Default = 250</li><li>Decimal Places Displayed = 2</li></ul>    |
  | Flowcell Type                    | Text Dropdown  | Required Field                                     | <p>Presets</p><ul><li>5M</li><li>25M</li><li>50M</li><li>100M</li></ul> |
  | Minimum Per Sample Volume (ul)   | Numeric        | Required Field                                     | <ul><li>Default = 2</li><li>Decimal Places Displayed = 2</li></ul>      |
* 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       |                                                    |                                                                                                        |
    | Derived Sample | Diluted Bulk Pool Volume (ul)    | Numeric        | Read Only                                          | Decimal Places Displayed = 1                                                                           |
    | Derived Sample | Diluted PhiX Volume (ul)         | Numeric        | Read Only                                          | Decimal Places Displayed = 1                                                                           |
    | Derived Sample | Final Loading Concentration (pM) | Numeric        | Required Field                                     | <ul><li><p>Presets</p><ul><li>225</li><li>400</li></ul></li><li>Decimal Places Displayed = 0</li></ul> |
    | Derived Sample | Final Loading Volume (ul)        | Numeric        | Read Only                                          | Decimal Places Displayed = 2                                                                           |
    | Derived Sample | KLD Volume (ul)                  | Numeric        | Read Only                                          | Decimal Places Displayed = 1                                                                           |
    | Derived Sample | MiSeq i100 Series Flowcell Type  | Text Dropdown  | <ul><li>Required Field</li><li>Read Only</li></ul> | <p>Presets</p><ul><li>5M</li><li>25M</li><li>50M</li><li>100M</li></ul>                                |
    | Derived Sample | RSB Volume (ul)                  | Numeric        | Read Only                                          | Decimal Places Displayed = 2                                                                           |
    | Derived Sample | Sample Name                      | Built-in       |                                                    |                                                                                                        |
    | Project        | Project Name                     | Built-in       |                                                    |                                                                                                        |

### Step 3: Load to Dry Cartridge (MiSeq i100 Series Sequencing v2.0)

* Master Step Name = Load to Dry Cartridge (MiSeq i100 Series Sequencing v2.0)
* Step Type = Standard
* Derived Sample Generation = Fixed, 1
* Naming Convention = {InputItemName}

{% hint style="info" %}
The version of Load to Dry Cartridge (MiSeq i100 Series Sequencing) master step name may be different depending on the version of IPP installed.
{% endhint %}

#### Automations

<details>

<summary>Validate Single Pool and Register Step Started</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon entry

{% code overflow="wrap" %}

```markup
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:validateSampleCount -min 1 -max 1 && \
/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_started.groovy'"
```

{% endcode %}

</details>

<details>

<summary>Validate Dry Cartridge Barcode</summary>

* Trigger Location = Placement
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
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:validate_output_containers -l {compoundOutputFileLuid1} -r 'MiSeq i100 Series Dry Cartridge:SC[0-9]{7}-SC3' -min 1 -max 1"
```

{% endcode %}

</details>

<details>

<summary>Validate Run Setup and Create Sample Sheet</summary>

* Trigger Location = Record Details
* Trigger Style = Manual button

{% code overflow="wrap" %}

```markup
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 \
          -exp 'if (!step.::Run Name::.matches(::[a-zA-Z0-9-_]+[a-zA-Z0-9-_. ]*::)) { fail(::Run Name contains prohibited characters. Please check to make sure Run Name only contains alphanumeric characters, spaces, dashes and underscores. Run Name must start with alphanumeric, a dash or an underscore::); }; if (step.::Run Name::.length() > 255) { fail(::Run Name shall not exceed 255 characters.::); };  if (step.::Index 2 Cycles:: > 0 && step.::Index 1 Cycles:: == 0) { fail(::Index 1 Cycles cannot be zero if Index 2 Cycles is non-zero::); };' \
          -log {compoundOutputFileLuid1} \
          -t true \
          && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/miseqi100series/automation/miseqi100series-automation.jar  script:generate_sample_sheet -i {stepURI:v2} -u {username} -p {password} -c {compoundOutputFileLuid0} -l {compoundOutputFileLuid1} -n 'INPUT.Instrument ID' --instrumentType '{udf:Instrument Type}' --verifyIndexLength"
```

{% endcode %}

</details>

<details>

<summary>Set Next Step</summary>

* Trigger Location = Record Details
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
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 {compoundOutputFileLuid1}"
```

{% endcode %}

</details>

<details>

<summary>Register Step Completed</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_completed.groovy'"
```

{% endcode %}

</details>

#### Queue

* 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 | <p>Instrument ID</p><blockquote><p>ℹ Only applicable for on-premise integration</p></blockquote> | Text           | Read Only                                          |                                                                         |
  | Derived Sample | MiSeq i100 Series Flowcell Type                                                                  | Text Dropdown  | <ul><li>Required Field</li><li>Read Only</li></ul> | <p>Presets</p><ul><li>5M</li><li>25M</li><li>50M</li><li>100M</li></ul> |
  | Derived Sample | Sample Name                                                                                      | Built-in       |                                                    |                                                                         |
  | Derived Sample | Waiting                                                                                          | Built-in       |                                                    |                                                                         |
  | Project        | Project Name                                                                                     | Built-in       |                                                    |                                                                         |

#### 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 | MiSeq i100 Series Flowcell Type | Text Dropdown  | <ul><li>Required Field</li><li>Read Only</li></ul> | <p>Presets</p><ul><li>5M</li><li>25M</li><li>50M</li><li>100M</li></ul> |
  | 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 = Row
* Destination Containers
  * MiSeq i100 Series Dry Cartridge
    * Rows
      * Number: 1
      * Naming: Alphabetic
    * Columns
      * Number: 1
      * Naming: Numeric
      * Start at: 1

#### Record Details

* Step Data (Master Step Fields)

  | **Field Name**                                                                                 | **Field Type** | **Options**    | **Additional Options and Dropdown Items**                              |
  | ---------------------------------------------------------------------------------------------- | -------------- | -------------- | ---------------------------------------------------------------------- |
  | Create Planned Run                                                                             | Toggle Switch  | Required Field | Default = True                                                         |
  | Index 1 Cycles                                                                                 | Numeric        | Required Field | <p>Presets</p><ul><li>0</li><li>6</li><li>8</li></ul>                  |
  | Index 2 Cycles                                                                                 | Numeric        | Required Field | <p>Presets</p><ul><li>0</li><li>6</li><li>8</li></ul>                  |
  | <p>Instrument</p><blockquote><p>ℹ Only applicable for on-premise integration.</p></blockquote> | Text           | Read Only      |                                                                        |
  | Read 1 Cycles                                                                                  | Numeric        | Required Field | <p>Presets</p><ul><li>51</li><li>151</li><li>301</li><li>501</li></ul> |
  | Read 2 Cycles                                                                                  | Numeric        | Required Field | <p>Presets</p><ul><li>51</li><li>151</li><li>301</li><li>501</li></ul> |
  | Run Name                                                                                       | Text           | Required Field |                                                                        |
* 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       |             |                                           |
    | Derived Sample | Sample Name         | Built-in       |             |                                           |
    | Project        | Project Name        | Built-in       |             |                                           |

### Step 4: AUTOMATED - Sequencing Run (MiSeq i100 Series Sequencing v2.0)

* Master Step Name = AUTOMATED - Sequencing Run (MiSeq i100 Series Sequencing v2.0)
* Step Type = Analysis
* Measurement Generation = Variable
* Naming Convention = Lane {OutputItemNumber}
* Reagent Kits
  * MiSeq i100 Series Dry Cartridge
    * Supplier = Illumina
  * MiSeq i100 Series Wet Cartridge
    * Supplier = Illumina

{% hint style="info" %}
The version of AUTOMATED - Sequencing Run (MiSeq i100 Series Sequencing) master step name may be different depending on the version of IPP installed.
{% endhint %}

#### Automations

<details>

<summary>Register Step Started and Register MiSeq i100 Series Run and Register MiSeq i100 Series Run Association and Register MiSeq i100 Series RunStarted</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon entry

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_started.groovy' && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/miseqi100series/register_miseqi100series_run.groovy' && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/miseqi100series/miseqi100series_associate_seqrun.groovy' && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/miseqi100series/miseqi100series_seqrun_started.groovy'"
```

{% endcode %}

</details>

<details>

<summary>Register MiSeq i100 Series RunCompleted and Register Step Completed</summary>

* Trigger Location = Step
* Trigger Style = Automatic upon exit

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/miseqi100series/miseqi100series_seqrun_completed.groovy' && /opt/gls/clarity/bin/java -jar /opt/gls/clarity/extensions/unified-product-analytics/automation/unified-product-analytics-automation.jar script:executeUPAAutomationScript -i {stepURI:v2} -u {username} -p {password} -s 'com/illumina/upa/scripts/common/step_completed.groovy'"
```

{% endcode %}

</details>

#### Queue

* 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       |             |                                           |
  | Derived Sample | Sample Name         | Built-in       |             |                                           |
  | Derived Sample | Waiting             | Built-in       |             |                                           |
  | Project        | Project Name        | Built-in       |             |                                           |

#### 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       |             |                                           |
  | 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** |
  | ----------------------------------- | -------------- | ----------- | ----------------------------------------- |
  | Current Cycle                       | Text           | Read Only   |                                           |
  | Current Read                        | Text           | Read Only   |                                           |
  | Dry Cartridge Barcode Serial Number | Text           | Read Only   |                                           |
  | Dry Cartridge Expiration Date       | Text           | Read Only   |                                           |
  | Flow Cell ID                        | Text           | Read Only   |                                           |
  | Flow Cell Type                      | Text           | Read Only   |                                           |
  | Instrument Control Software Version | Text           | Read Only   |                                           |
  | Instrument ID                       | Text           | Read Only   |                                           |
  | Instrument Type                     | Text           | Read Only   |                                           |
  | Output Folder                       | Text           | Read Only   |                                           |
  | Run Name                            | Text           | Read Only   |                                           |
  | Run Status                          | Text           | Read Only   |                                           |
  | Sequencing Log                      | Multiline Text | Read Only   |                                           |
  | Wet Cartridge Expiration Date       | Text           | Read Only   |                                           |
  | Wet Cartridge Serial Number         | Text           | Read Only   |                                           |
* 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      | % Aligned R1         | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Aligned R2         | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Bases >=Q30 R1     | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Bases >=Q30 R2     | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Error Rate R1      | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Error Rate R2      | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Occupied           | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % PF                 | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | % Phasing R1         | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Container      | % Phasing R2         | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Container      | % Prephasing R1      | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Container      | % Prephasing R2      | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Container      | Container Name       | Built-in       |             |                                           |
    | Container      | Intensity Cycle 1 R1 | Numeric        | Read Only   |                                           |
    | Container      | Intensity Cycle 1 R2 | Numeric        | Read Only   |                                           |
    | Container      | LIMS ID (Container)  | Built-in       |             |                                           |
    | Container      | Reads PF             | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | Yield (Gb) R1        | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Container      | Yield (Gb) R2        | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Derived Sample | Sample Name          | Built-in       |             |                                           |
    | Measurement    | % Aligned R1         | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | % Aligned R2         | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | % Bases >=Q30 R1     | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | % Bases >=Q30 R2     | Numeric        | Read Only   |                                           |
    | Measurement    | % Error Rate R1      | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | % Error Rate R2      | Numeric        | Read Only   |                                           |
    | Measurement    | % Occupied           | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | % PF                 | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | % Phasing R1         | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Measurement    | % Phasing R2         | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Measurement    | % Prephasing R1      | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Measurement    | % Prephasing R2      | Numeric        | Read Only   | Decimal Places Displayed = 3              |
    | Measurement    | Intensity Cycle 1 R1 | Numeric        | Read Only   |                                           |
    | Measurement    | Intensity Cycle 1 R2 | Numeric        | Read Only   |                                           |
    | Measurement    | Reads PF             | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | Yield (Gb) R1        | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Measurement    | Yield (Gb) R2        | Numeric        | Read Only   | Decimal Places Displayed = 2              |
    | Project        | Project Name         | Built-in       |             |                                           |

### Step 5: AUTOMATED - Analysis Run (MiSeq i100 Series Sequencing v2.0)

* Master Step Name = AUTOMATED - Analysis Run (MiSeq i100 Series Sequencing v2.0)
* Step Type = Demultiplexing
* Measurement Generation = Fixed, 1
* Naming Convention = {SubmittedSampleName} {AppliedReagentLabels}

{% hint style="info" %}
The version of AUTOMATED - Analysis Run (MiSeq i100 Series Sequencing) master step name may be different depending on the version of IPP installed.
{% endhint %}

#### Queue

* 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       |             |                                           |
  | Derived Sample | Sample Name         | Built-in       |             |                                           |
  | Derived Sample | Waiting             | Built-in       |             |                                           |
  | Project        | Project Name        | Built-in       |             |                                           |

#### 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       |             |                                           |
  | 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** |
  | ------------------------ | -------------- | ----------- | ----------------------------------------- |
  | Analysis Result Location | Text           | Read Only   |                                           |
  | Analysis Status          | Text           | Read Only   |                                           |
  | Log                      | Multiline Text | Read Only   |                                           |
* 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       |             |                                           |
    | Derived Sample | Sample Name         | Built-in       |             |                                           |
    | Project        | Project Name        | Built-in       |             |                                           |


---

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