The integration includes the following features:
Preconfigured NovaSeq X Series Sequencing v1.0 workflow that maps to lab protocols and instrument runs.
The following steps in the NovaSeq X Series Sequencing v1.0 preconfigured protocol:
Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0)
Make Bulk Pool (NovaSeq X Series Sequencing v1.0)
Dilute and Denature (NovaSeq X Series Sequencing v1.0)
Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0)
AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0)
Automatic validation of run setup information before sample sheet generation or planned run creation.
Automated generation of the sample sheet. This sample sheet is used to start the sequencing run on the NovaSeq X Series Control Software through Local run mode.
Automated creation of a planned run on Illumina Connected Analytics (ICA). The NovaSeq X Series Control Software retrieves the planned run for use with the Cloud run mode.
Automated tracking of the NovaSeq X Series sequencing run and parsing of run statistics into Clarity LIMS.
Support for configuration of multiple secondary analyses with multiple library prep kits (LPKs) and index adapter kits (IAKs) for the planned run. The analysis is either performed by ICA in Cloud mode or by DRAGEN on-board the instrument in Local mode.
[Optional] Preconfigured Library Prep Validation v2.3.1 workflow used for validation purposes only. The workflow contains a single-step protocol that models the library prep required to produce libraries that are ready for the NovaSeq X Series Sequencing v1.0 workflow. For more information, refer to NovaSeq X Series Integration v1.1.0 User Interaction, Validation and Troubleshooting.
[Optional] Clarity LIMS Product Analytics (CLPA) integration that can be used with ICA. The CLPA integration installation and configuration is required. For more information, refer to Clarity LIMS Product Analytics Integration v1.2 Configuration.
Last Updated: November 2024
Release Date: September 2023
Document Version: 2
These release notes describe the key changes to software components for the Clarity LIMS NovaSeq X Series Integration Package v1.1.0.
Refer to Compatibility under Instruments & Integrations.
Updated Java and third-party dependency libraries.
Updated Groovy to v3.0.7.
Restricted the scope for the JWT token used in the integration.
Fixed issue where the SIS core service was not properly registered in systemctl.
This integration does not track the analysis step. You must log in to BSSH to view the cloud analysis status and results. For local analysis, the secondary analysis results are located in the external storage configured in Illumina Run Manager.
The Local run mode does not support custom library prep and index adapter kits.
The Analysis Configuration Template (ACT) names must be unique.
The Assign Analysis Configuration Template step expects the input samples to be unpooled libraries.
On the Make Bulk Pool step, the log displays a warning when the Calculate Volume automation is triggered and at least one pool consists of multiple inputs. This issue is caused by the output custom field being reset multiple times at the end of the automation. This issue does not affect the Calculate Volume automation functionality.
Sample sheet and planned run generation will fail if any of the samples in the pools has been assigned QC flag in prior steps before entering the Load to Library Tube Strip step.
Version
Changes
2
Updated Compatibility section to reference Compatibility matrix table.
Updated Known Limitations section.
1
Initial release.
This section explains the user interaction for each step and how to validate the installation of the Illumina NovaSeq X Series Integration Package v1.1.0.
The validation process involves the following actions:
Running samples through the Library Prep Validation v2.3.1 workflow.
The workflow contains a single-step protocol that models the library prep required to produce libraries tagged with index sequences. At the end of the step, these libraries are routed to NovaSeq X Series Sequencing v1.0 workflow.
Running libraries through the NovaSeq X Series Sequencing v1.0 workflow validates the following items:
Successful sequential step advancement of samples through the steps of the workflow.
Automatic validation of run setup information before sample sheet generation or planned run creation.
Automated sample sheet generation. This sample sheet is used to start the sequencing run on the NovaSeq X Series Control Software through the Local run mode.
Automated creation of a planned run in Illumina Connected Analytics (ICA). The control software retrieves the planned run. The run is started through the Cloud run mode.
Automated tracking of the NovaSeq X Series sequencing run and parsing of run statistics into Clarity LIMS.
The validation steps assume that the following conditions have been met:
NovaSeq X Series Integration Package v1.1.0 is installed and you have imported the default Clarity LIMS configuration.
Analysis configuration templates (ACTs) are created in BaseSpace Sequence Hub for your run. Make sure that the index adapter kit label group is created in Clarity LIMS before selecting it in the ACT. This same label group is used in the Run Library Prep Validation v2.3.1 step. For more information on creating a reagent label group, refer to Add and Configure Labels and Label Groups in the Clarity LIMS (Clarity & LabLink Reference Guide) documentation. For the adapter sequences for your library prep kits, refer to Illumina Adapter Sequences.
You must create the Analysis Configuration Templates (ACTs) that are required for configuring secondary analysis in the NovaSeq X Series Sequencing v1.0 workflows. Create and delete ACTs in BaseSpace Sequence Hub. For instructions, refer to the BaseSpace Sequence Hub Online Help on the Illumina support site.
The following steps set up Clarity LIMS in preparation for running samples through the Library Prep Validation v2.3.1 and NovaSeq X Series Sequencing v1.0 workflows.
On the Configuration tab, under Workflows, activate both the Library Prep Validation v2.3.1 and NovaSeq X Series Sequencing v1.0 workflows.
On the Projects and Samples screen, create a project and add samples to it.
Assign the samples to the Library Prep Validation workflow.
This single-step protocol models the library prep required to produce libraries tagged with index sequences that are ready for the NovaSeq X Series Sequencing v1.0 workflow.
Follow the steps in Library Prep Validation Protocol to run the Library Prep Validation workflow with the following:
Label Group = same as the Index Adapter Kit selected in the ACT that is being used
Sequencing Instrument = NovaSeq X Series
On exit from the step, the Routing Script automation is triggered. This automation assigns samples to the first step of the NovaSeq X Series Sequencing v1.0 workflow, Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0).
The NovaSeq X Series Sequencing v1.0 protocol consists of the following steps:
Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0)
Make Bulk Pool (NovaSeq X Series Sequencing v1.0)
Dilute and Denature (NovaSeq X Series Sequencing v1.0)
Load To Library Tube Strip (NovaSeq X Series Sequencing v1.0)
AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0)
Each step contains a script to register the start time (upon step entry) and the time that the step is completed (upon step exit). This information is published to CLPA through ICA. These scripts are used only for CLPA support and can be incorporated as part of an automation that performs other functions.
In Lab View, locate the NovaSeq X Series Sequencing v1.0 protocol. The samples are queued for the Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0) step.
Add the samples to the Ice Bucket and select View Ice Bucket.
On the Ice Bucket screen, select Begin Work to start the Validate Sample Names and Retrieve Analysis Configuration Template List and Register Step Started automation.
The automation checks that the sample names do not use restricted characters and are within character limits and retrieves the list of ACTs that you can select. The automation also registers the start time of the step by publishing messages to CLPA through ICA. The part of the script for start time registration is used only for CLPA support.
Select the applicable ACT to assign to the samples. The index adapter kit specified by the ACT must correspond with the label group used in Library Prep Protocol.
[Optional] Under Step Details, select Retrieve ACT Information to trigger the Retrieve ACT Information automation.
This automation retrieves ACT information (e.g., Library Prep Kit, Index Adapter Kit, Reference Genome, and so on) and populates the fields in Clarity LIMS. These details are saved to the ACTMetadata.csv file that you can download. If you are not sure of the analysis configuration before starting the sequencing and analysis run, refer to the details in the file.
Select Next Steps to assign the ACT to the samples.
This action triggers the Validate Reagent Labels and Apply Selected ACT to Samples and Set Next Step automation. This automation validates that the indexes applied to the libraries are valid for the selected ACT and assigns samples to it. All samples added to the Ice Bucket (mentioned above) are assigned to this ACT.
Select Finish Step.
To use multiple ACTs in a single run, repeat 2–7 for each ACT. For step 2, only add samples that are being assigned to that ACT into the Ice Bucket.
In Lab View, locate the NovaSeq X Series Sequencing v1.0 protocol. The samples are queued for the Make Bulk Pool (NovaSeq X Series Sequencing v1.0) step.
Add the samples to the Ice Bucket and select View Ice Bucket.
You can also add control samples by selecting them from the Available control samples drop-down list and selecting Add.
On the Ice Bucket screen, select Begin Work.
Create a pool of samples as follows.
On the Pooling screen, create a pool by dragging samples into the Pool Creator.
If a control sample was added, it appears under the Sample List and can be pulled into the pool that needs the control.
Enter a name for your pool or accept the default name (Pool #1).
[Optional] If multiple pools are required, select the plus sign (+) next to Pool Creator to create a pool.
[Optional] To remove a pool, select the X in the top right corner of the pool.
Select Record Details to trigger the Validate Analysis Configurations automation.
This automation performs the following checks on the analysis configuration for each pool:
Pooled samples are within the maximum configuration limit.
Pooled samples have the sample type of analysis (Cloud or Local).
Pooled samples that have the same secondary analysis also have the same analysis version and settings.
On the Record Details screen, navigate to the Reagent Lot Tracking section to track the lot information used in the step.
[Optional] Create a new lot. For more information, refer to Add and Configure Reagent Kits and Lots in the Clarity LIMS (Clarity & LabLink Reference Guide) documentation.
In the Step Details area, complete the following required fields:
Number of Lanes to Sequence — Used in volume calculations, to make sure that the volumes are sufficient for the number of times the pool is sequenced. This field is applied to all pools in the step.
Final Loading Concentration (pM) — The final loading concentration of the pool in the flow cell.
Final Loading Volume (ul) — The final loading volume of the pool in the flow cell. This field is prepopulated with the configured default value (200 ul), but can be edited if more volume is required.
Minimum Per Sample Volume (ul) — The minimum volume for each sample. This field is prepopulated with the configured default value (2 ul) and can be edited. If the per sample volume is below the value set in this field, the Calculate Volumes script applies a rounding factor to affected samples so that the volume reaches the minimum volume.
% PhiX (2.0nM) Spike-In — The value entered in this field is used to calculate the volume of PhiX spike-in required.
Flowcell Type — This is a read-only field configured with the default value of B3.
Select Calculate Volumes to trigger the Calculate Volumes automation.
This automation calculates the volumes required for each library to form a pool that has the concentration and volume specified in the Step Details fields. The automation is applied to control samples when they are indexed and molarity is specified. It also generates the calculation file in a CSV format which is available for download under Files placeholder section below.
In the Sample Details table, select the pool next to the sample name to view details on the pool composition.
Select Next Steps to trigger the Set Next Step automation.
This automation sets the next step for samples to ADVANCE, which moves them to the Dilute and Denature (NovaSeq X Series Sequencing v1.0) step.
Select Finish Step.
In Lab View, locate the NovaSeq X Series Sequencing v1.0 protocol. The pool of samples queued for the Dilute and Denature (NovaSeq X Series Sequencing v1.0) displays.
Add the pool to the Ice Bucket and select View Ice Bucket.
[Optional] On the Ice Bucket screen, set the number of derivatives to create (placed into the library tube strip) and select Begin Work.
On entry to the step, the Validate Inputs Flowcell Type and Register Step Started automation is triggered. This automation makes sure that the configured flow cell type is valid. The automation also registers the start time of the step by publishing messages to CLPA through ICA. The portion of the script used for start time registration is used only for CLPA support.
On entry to the Record Details screen, the Calculate Volumes automation is triggered. This automation sets the following values:
BP Aliquot Volume (ul)
RSB Volume (ul)
NaOH Volume (ul)
TT2 Volume (ul)
The automation also generates the calculation file (CSV) and attaches it to the step. This file contains information about the volume of RSB, NaOH, and TT2 to add per working pool.
On the Record Details screen, the Reagent Lot Tracking section tracks the NaOH, Resuspension Buffer, and TT2 reagents used in the step. To add and activate reagent lots, refer to Add and Configure Reagent Kits and Lots in the Clarity LIMS (Clarity & LabLink Reference Guide) documentation.
On the Record Details screen, perform the following actions:
In the Reagent Lot Tracking section, select from the active lots displayed in each drop-down list.
In the Sample Details table, the BP aliquot, RSB, NaOH, and TT2 reagent volume values are auto populated and read-only.
[Optional] In the Sample Details table, select the pool icon to view details of the working pool composition.
The working pool number is appended to the bulk pool name so that you can identify which working pools are derived from the same bulk pool.
In the Files area, download the Calculation File (CSV) and open it and view details on the RSB, NaOH, and TT2 volumes to add per working pool.
Select Next Steps.
Select Finish Step.
In Lab View, locate the NovaSeq X Series Sequencing v1.0 protocol. The pool of samples are queued for the Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) step.
Add the pools to the Ice Bucket and select View Ice Bucket.
On the Ice Bucket screen, select Begin Work to trigger the Validate Analysis Configurations and Register Step Started automation.
Validate Analysis Configurations automation performs the following basic checks on the analysis configuration for each pool:
Pooled samples are within the maximum configuration limit.
Pooled samples have the sample type of analysis (Cloud or Local).
Pooled samples that have the same secondary analysis also have the same analysis version and settings.
Register Step Started automation registers the start time of the step by publishing messages to CLPA through ICA. The portion of the script used for start time registration is used only for CLPA support.
On the Placement screen, do as follows.
Drag the pool into the Library 8-tube Strip in the Placed Samples area on the right.
Scan or type the barcode of the Library 8-tube Strip into the Library 8-tube Strip container name field.
Select Record Details.
After exiting the Placement screen, the Validate Library Strip Tube Barcode automation makes sure that the library tube strip barcode conforms to the barcode mask LC[0-9]{7}-LC1.
The fields displayed on the Record Details screen are used to create the planned run and generate the sample sheet.
The analysis related information for the planned run is from the ACT associated with the samples and no further analysis configuration is required. Refer to the following table for details.
Fields Displayed on Record Details Screen of Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) Step
¹ The Run/Analysis configuration is imported manually to the NovaSeq X Series instrument through a sample sheet. The sample sheet generated on this step is downloaded and imported manually by the user on NovaSeq X Control Software to start the run. The downstream secondary analysis is done using the onboard DRAGEN module.
² The planned run configuration is downloaded to the NovaSeq X Series Instrument Control Software to start the run. The downstream secondary analysis is done on the cloud.
³ The custom value must correspond to the longest index sequence of the samples in the pools in the library tube strip. Otherwise, the planned run creation fails and an error message displays.
Select Validate Run Setup and Create Planned Run to trigger the automation script. The script performs the following actions:
Validates the parameters entered on the Record Details screen.
Clarity consolidates sample/library information, run and analysis configuration details and send them to ICA to create a planned run and samplesheet for Cloud run mode.
Clarity consolidates sample/library information, run and analysis configuration details and send them to ICA to generate samplesheet for Local run mode.
Select Next Steps.
On the Assign Next Steps screen, the next step for the pooled samples is set to the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step.
Select Finish Step to advance the pooled samples to the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step. For more information on how to start the sequencing run for different run modes, refer to NovaSeq X Series Integration v1.1.0 Configuration.
To use multiple ACTs in a single run, repeat steps 1–3 for each ACT. The pool of samples (for different ACTs) are then queued for the Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) step to be added to the Ice Bucket.
This protocol contains the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step. This step is fully automated.
The integration starts the step automatically and data from the run is parsed back into Clarity LIMS. User interaction is not required, but you can review the stages of the step in Clarity LIMS.
Read summary metrics are recorded for the library pool. After the run is complete, open the step and review these metrics in the Step Details section and the Sample Details table.
If an automation trigger does not appear to run its corresponding scripts, refer to the Troubleshooting Automation section in the Clarity LIMS (API & Database) documentation.
If the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step does not start, Clarity LIMS is likely not receiving sequencing run events from BaseSpace Sequence Hub correctly. Check for the following issues:
Make sure that the NovaSeq X Control Software is configured properly as follows.
Before you start the run, in the NovaSeq X Control Software, select the appropriate BaseSpace Sequence Hub region in the Hosting location drop-down list.
Under Run Settings, check the boxes next to Cloud run monitoring and Cloud run storage.
If there are issues after modifying the configuration settings on the NovaSeq X Control Software, contact Illumina Support.
If the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step starts, but does not complete, do as follows.
Log in to Clarity LIMS and use one of the following methods to open the in-progress step:
In Lab View, find the step in the Recent Activities pane.
Search for the step in Clarity LIMS using the library tube strip barcode as the search term.
On the Record Details screen, the Sequencing Log multiline text field contains logging information.
If unable to reach the Record Details screen, or if the Sequencing Log field does not contain enough information to resolve the issue, contact Illumina Support. Provide the relevant information from the troubleshooting steps already performed.
The Validate Analysis Configuration automation check is in the Make Bulk Pool (upon pooling) and Load to Library Tube Strip (upon step entry) steps. If a failure happens, an error message displays and the step can be aborted, or you might be prevented from continuing the step. This check can fail for the following reasons:
The analysis configuration after library pooling or during the planned run creation exceeds the configuration limit.
Resolve this error as follows.
If the error occurs on the Make Bulk Pool step, reduce the number of analysis configurations for a pool. To reduce the number, reorganize the samples for the pooling step (eg, by pooling samples that have similar configurations together).
If the error occurs on the Load to Library Tube step, reduce the number of analysis configurations for the planned run by reorganizing the samples for multiple runs instead of a single run.
ACTs of samples in the same pool or planned run must have the same run mode (Local or Cloud).
Resolve this error as follows.
Abort the step and remove samples with ACTs that have conflicting run modes from the Ice Bucket.
Make sure that all remaining samples in the Ice Bucket have ACTs with the same run modes.
Select Begin Work to continue the step.
ACTs of samples in the same pool or planned run that have the same secondary analysis must have the same analysis version.
Resolve this error as follows.
If the error occurs on the Make Bulk Pool step, pool the samples again. Make sure that samples with the same secondary analysis, but conflicting analysis versions, are in different pools.
If the error occurs on the Load to Library Tube step, reorganize the samples for the planned run. Make sure that samples with the same secondary analysis, but conflicting analysis versions, are in different runs.
ACTs of samples in the same pool or planned run that have the same secondary analysis must have the same analysis settings. The analysis setting fields can differ for a different secondary analysis and are configured in BaseSpace Sequence Hub when the ACT is created.
Resolve this error as follows.
If the error occurs on the Make Bulk Pool step, pool the samples again. Make sure that samples with the same secondary analysis, but conflicting analysis settings, are in different pools.
If the error occurs on the Load to Library Tube step, reorganize the samples for the planned run. Make sure that samples with the same secondary analysis, but conflicting analysis settings, are in different runs.
The Illumina NovaSeq X Series Integration Package v1.1.0 supports the integration of Clarity LIMS to Illumina NovaSeq X series instruments.
For user interactions for each step and instructions on validating and troubleshooting the Illumina NovaSeq X Series Integration, refer to .
The configuration provided in this integration has been established to support NovaSeq X Series lab processes. Any configuration changes to protocols or workflows (including renaming protocols, steps, and fields) could break the process.
The Illumina Universal Sample Identifier is a global field that is imported with the Library Prep Validation v2.3.1 and NovaSeq X Series Sequencing v1.0 workflows. Illumina Universal Sample Identifier is a text field that is reserved for CLPA support. The field is optional and not required for this integration.
The NovaSeq X Series Sequencing v1.0 workflow uses analysis configuration templates (ACTs) to configure secondary analysis for planned runs. Required ACTs must be created using BSSH. The ACT names must be unique. The index adapter kit (the label group in Clarity LIMS) selected in the ACT is created in Clarity LIMS. The same label group must be used in the library preparation step. For more information, refer to .
Make sure that ACT names created in BaseSpace Sequence Hub do not have leading or trailing spaces. Otherwise, Clarity LIMS can have issues with recognizing the ACT names.
Samples must go through the library preparation and quantification process before they enter the NovaSeq X Series Sequencing v1.0 workflow. It is assumed that the following steps have completed before samples are assigned to the workflow:
Samples have been accessioned into Clarity LIMS.
Samples have been run through QC and library prep.
Samples have the Molarity (nM) global field set to a value. This value is required for the Calculate Volumes automation in the Make Bulk Pool step. For more information on sample accessioning, refer to Sample Accessioning and Upload and Modify Samples in the Getting Started section of the .
You can assign samples to workflows automatically, using a routing script, or manually—from the Projects & Samples dashboard. Refer to Assign and Process Samples in the .
The NovaSeq X Series Integration Package v1.1.0 includes the following workflows:
Library Prep Validation v2.3.1 (optional, but recommended for validation purposes)
NovaSeq X Series Sequencing v1.0
The following describes the protocols and steps included in these workflows.
The following automations are configured on this step:
Validate Sample Names and Retrieve Analysis Configuration Template List and Register Step Started²
Retrieve ACT Information
Validate Reagent Labels and Apply Selected ACT to Samples and Set Next Step
Register Step Completed¹
¹ These automations are required for CLPA support only.
² These automations are required for the NovaSeq X Series Sequencing v1.0 workflows and contain additional logic needed for CLPA support. If you would like to remove support for CLPA, contact Illumina Support.
All remaining automations are required for the NovaSeq X Series Sequencing v1.0 workflows.
The following table lists the configuration details for fields that are defined on the Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0) step.
Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0) Master Step Field Configuration
The following table lists the global custom fields that are configured to display on the Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0) step.
Global Field Configuration (Configured on Derived Sample)
Samples are pooled in the Make Bulk Pool (NovaSeq X Series Sequencing v1.0) step. You can manually create working pools based on the final loading concentration required.
The following automations are configured on this step:
Register Step Started¹
Validate Analysis Configurations
Calculate Volumes
Set Next Step
Register Pools & Register Step Completed¹
¹ These automations are required for CLPA support only.
All remaining automations are required for the NovaSeq X Series Sequencing v1.0 workflows.
The following table lists configuration details for the fields that are defined on the Make Bulk Pool (NovaSeq X Series Sequencing v1.0) step.
Make Bulk Pool (NovaSeq X Series Sequencing v1.0) Master Step Field Configuration
The following table lists the global custom fields that are configured to display on the Make Bulk Pool (NovaSeq X Series Sequencing v1.0) step.
Global Custom Fields Configuration (Configured on Derived Sample)
The Dilute and Denature (NovaSeq X Series Sequencing v1.0) step allows you to dilute pooled samples with the addition of RSB.
The following automations are configured on this step:
Validate Inputs Flowcell Type and Register Step Started²
Calculate Volumes
Set Next Step
Register Step Completed¹
¹ These automations are required for CLPA support only.
² These automations are required for the NovaSeq X Series Sequencing v1.0 workflows and contain additional logic needed for CLPA support. If you would like to remove support for CLPA, contact Illumina Support.
All remaining automations are required for the NovaSeq X Series Sequencing v1.0 workflows.
The following table lists the global custom fields that are configured to display on the Dilute and Denature (NovaSeq X Series Sequencing v1.0) step.
Global Field Configuration (Configured on Derived Sample)
The Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) step allows you to scan the library tube strip barcode into Clarity LIMS. Then, you can place the working pools into the library tube strip used in the NovaSeq X Series run. This step also does the following actions:
Validates the run setup and analysis information.
Generates the sample sheet file.
Creates a planned run on ICA, depending on the selected run mode.
¹ These automations are required for the NovaSeq X Series Sequencing v1.0 workflows and contain additional logic needed for CLPA support. If you would like to remove support for CLPA, contact Illumina Support.
² These automations are required for CLPA support only.
All remaining automations are required for the NovaSeq X Series Sequencing v1.0 workflows.
The following table shows the master step fields that are configured on the Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) step. These fields are required for sample sheet generation and planned run creation in ICA.
Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) Master Step Field Configurationn
The AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step is fully automated. The integration starts the step, adds samples to the Ice Bucket, and completes the step. Data from the run is parsed back to Clarity LIMS. In this step, pooled samples in the library tube strip are sequenced on the NovaSeq X Series instrument.
¹ These automations are required for CLPA support only.
All remaining automations are required for the NovaSeq X Series Sequencing v1.0 workflows.
The following tables show the master step fields that are configured on the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step.
Clarity LIMS Master Step Field Information
Other Clarity LIMS Master Step Fields
The following global custom fields are used to capture the run metrics in Clarity LIMS:
% Bases >=Q30 R1
% Bases >=Q30 R2
% Error Rate R1
% Error Rate R2
Yield (Gb) R1
Yield (Gb) R2
Reads PF R1
Reads PF R2
%PF R1
%PF R2
% Aligned R1
% Aligned R2
% Phasing R1
% Phasing R2
% Prephasing R1
% Prephasing R2
Intensity Cycle 1 R1
Intensity Cycle 1 R2
Cluster Density R1
Cluster Density R2
At the end of the step, the pools of samples are automatically removed from the step. The step completes automatically when Run Status is RunCompletedSuccessfully.
The following information summarizes how the NovaSeq X Series integration works.
After the Validate Run Setup and Create Planned Run automation is triggered on the Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) step, the run parameters entered in the Run Details screen are sent to ICA. The analysis configuration parameters from the selected ACTs are also sent to ICA. ICA validates the run and analysis configuration. If the validation fails, ICA sends an error message to Clarity LIMS. If the validation passes, ICA generates the sample sheet and sends it back to Clarity LIMS. ICA also creates the planned run based on the selected run mode (eg, Local or Cloud).
For the Local run mode, the sample sheet is generated and stored in Clarity LIMS. This sample sheet contains the run and analysis configuration information required to start the run on the NovaSeq X Series instrument.
For the Cloud run mode, the planned run is created in ICA. The planned run contains the run and analysis configuration information required to start the run on the instrument. The sample sheet is generated and stored in Clarity LIMS for reference purposes.
When the sequencing run starts on the instrument, the NovaSeq X Series Control Software notifies BSSH. BSSH monitors the run and uses the Stratus Integration Service (SIS) to notify Clarity LIMS. The events are processed and the integration service retrieves the run information from BSSH. This information is used to populate the custom fields in the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step.
Other run events follow the same information flow. When sequencing is complete, the control software uploads the sequencing run data (primary metrics) and associated files to ICA. Then, Clarity LIMS retrieves the primary metrics and uses them to populate the fields in the Sample Details table (eg, % Error Rate R1). The custom fields (eg, Run Status, Current Read, and so on) on the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step are updated using the run related information. If the sequencing run is successfully completed, the step automatically completes.
The analysis results are not tracked. You must log in to BSSH to retrieve the cloud analysis results. For local analysis, the secondary analysis results are found in the external storage configured in Illumina Run Manager. The external storage information is found in the External Storage for Analysis Results configuration settings in Illumina Run Manager.
The following sections outline the steps to start a Clarity LIMS cloud or local run on the NovaSeq X Series instrument.
You can start the sequencing run in BSSH for both the Local and Cloud modes. Make sure that you log in using the account that was used during the NovaSeq X Series integration package setup. You must also import the sample sheet created from the Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0) step for the Local Run Mode.
The following sections describe the components (files, properties, reagent categories / label groups, reagent kits, and containers) that are installed by default as part of this integration.
Reagent Kits
Buffer Cartridge
Lyophilization Cartridge
NaOH
Reagent Cartridge
Resuspension Buffer (RSB)
TT2
Container Types
Library 8-Tube Strip
Tube This integration supports the library 8-tube strip with the barcode in the LC[0-9]{7}-LC1 format (eg, LC1234567-LC1).
Control Types
PhiX v3
The workflow configuration contains several validation checks. To make sure that the calculations work properly, it is important that you do not disable any of this validation logic. The validation checks determine the following information:
Which samples, and how many, can enter each step together.
Which samples, and how many, can be pooled together.
All submitted samples must have an associated secondary analysis that is configured using the analysis configuration template (ACT). The ACT must be configured on BSSH before starting the Assign Analysis Configuration Template step. The ACT names must be unique.
The library tube strip barcode must be unique. There must not be multiple library tube strip containers with the same name in the system.
Reagent labels, or indexes, must be unique.
One library pool can only contain one library or control with no label/index.
The AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step must not be manually started or completed. This step is fully automated and the sequencing service does not update samples correctly if they have been manually started.
For the automated run to start successfully, you must select Validate Run Setup and Create Planned Run in the Load to Library Tube step.
Only use alphanumeric, dash, or underscore characters in the submitted sample names. Invalid characters can cause a sample sheet validation failure in the Load to Library Tube Strip step.
Do not add samples to the Ice Bucket or start or complete the AUTOMATED - Sequencing Run (NovaSeq X Series Sequencing v1.0) step. The integration does this automatically.
The Library Prep Validation v2.3.1 workflow allows for validation of the system after installation is complete. For details, refer to .
The Assign Analysis Configuration Template (NovaSeq X Series Sequencing v1.0) step uses ACTs to configure secondary analysis for samples. Each ACT contains details related to the secondary analysis (eg, index adapter kit to use, reference genome setting, and so on). You can select an ACT from a list and assign samples to it. A planned run can have multiple ACTs and this step must be repeated for each ACT that is required for the run. For more information on creating ACTs and assigning samples to them, refer to .
For more information on sample name character restrictions, refer to .
If a planned run with the same sample name and project name (case-insensitive) has been created previously in ICA, the sample sheet generated from the Validate Run Setup and Create Planned Run automation can reflect the original case of the previous sample name. This can cause validation errors for analysis configurations with sample-level settings. To resolve this issue, change the sample name or the project name on Clarity LIMS and run the automation again.
. Select Cloud run storage options under Run Settings.
Follow instructions to start a sequencing run with local secondary analysis in .
Follow instructions to start a sequencing run with cloud secondary analysis in .
The library preparation workflow of the samples must be before routing the samples through the library preparation workflow.
Field
Value
Run Name
Enter the experiment name. Only alphanumeric characters, dashes, and underscores are permitted. No spaces.
Run Mode
The selected Run Mode must correspond to the ACT type. Presets
Local¹
Cloud²
Read 1 Cycles
Presets
151
101
51
Type a custom value
Read 2 Cycles
Presets
151
101
51
Type a custom value
Index 1 Cycles
Presets
0
6
8
Type a custom value³
Index 2 Cycles
Presets
0
6
8
Type a custom value³
Field Name | Field Type | Options |
Analysis Configuration Template | Text Dropdown | Required Field |
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 |
Field Name | Field Type | Options | Additional Options and Dropdown Items |
Molarity (nM) | Numeric | Decimal places displayed = 2 |
ACT Name | Text | Read Only |
Field Name | Field Type | Options | Additional Options and Dropdown Items |
Final Loading Concentration (pM) | Numeric Dropdown |
|
|
RSB Volume (ul) | Numeric |
|
|
NovaSeq X Flowcell Type | Text |
|
|
Field Name | Field Type | Options | Additional Options and Dropdown Items |
BP Aliquot Volume (ul) | Numeric | Read Only | Decimal places displayed = 0 |
NaOH Volume (ul) | Numeric | Read Only | Decimal places displayed = 2 |
RSB Volume (ul) | Numeric | Read Only | Decimal places displayed = 2 |
TT2 Volume (ul) | Numeric | Read Only | Decimal places displayed = 2 |
Field Name | Source File | Corresponding Field in Source File | Additional Information |
Current Cycle |
|
|
|
Current Read |
|
|
|
Flow Cell Expiration Date |
|
|
|
Flow Cell ID |
|
|
|
Flow Cell Lot Number |
|
|
|
Flow Cell Part Number |
|
|
|
Flow Cell Side |
|
|
|
Flow Cell Type |
|
|
|
Instrument Control Software Version |
|
|
|
Instrument ID |
|
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|
Instrument Type |
|
|
|
Output Folder |
|
|
|
Library Tube Barcode |
|
|
|
Library Tube Lot Number |
|
|
|
Run End Time |
|
|
|
Run Name |
|
|
|
Field Name | Description | Additional Information |
Run Status |
| Visible on Record Details screen. |
Sequencing Log |
| Visible on Record Details screen. |
BaseSpace Run ID |
| Hidden |
ICA Project ID |
| Hidden |
Instrument Run ID |
| Hidden |
Run Start Time |
| Hidden |
Field Name | Field Type | Options | Additional Options and Dropdown Items |
% PhiX (2.5 nM) Spike-In | Numeric |
|
Final Loading Concentration (pM) | Numeric Dropdown |
|
Final Loading Volume (ul) | Numeric |
|
|
Flowcell Type | Text |
|
|
Minimum Per Sample Volume (ul) | Numeric |
|
|
Number of Lanes to Sequence | Numeric |
|
|
PhiX Volume (ul) | Numeric |
|
|
Field Name | Field Type | Options | Additional Options and Dropdown Items |
Cloud Run ID | Text | Read Only |
|
Index 1 Cycles | Numeric Dropdown |
|
|
Index 2 Cycles | Numeric Dropdown |
|
|
Output Folder | Text |
Read 1 Cycles | Numeric Dropdown |
|
|
Read 2 Cycles | Numeric Dropdown |
|
|
Run Mode | Text Dropdown | Required Field | Presets
|
Run Name | Text | Required Field |
Do not remove, rename, or modify this field as it can cause the integration to break.
Do not remove, rename, or modify this field as it can cause the integration to break.
