The Illumina NovaSeq 6000 Integration v2.5.0 supports the integration between Clarity LIMS and the NovaSeq 6000 instrument. This integration also provides support for Clarity LIMS v6.2.0 and includes updates to the technology stack and third party libraries and utilities.
Clarity LIMS is automatically stopped during installation of the RPM and will need to be restarted. The file-based and API-based integrations cannot run at the same time. When the API-based integration has finished installing, shut down Clarity LIMS and uninstall the file-based integration.
This document provides instructions for installing NovaSeq 6000 Integration v2.5.0. It also describes the components that are installed in the default configuration.
For details on installed protocols and steps, automations, generated and captured files, and rules and constraints, refer to NovaSeq 6000 Integration v2.5.0 Configuration.
For information on user interaction for each step, validating and troubleshooting the integration, refer to NovaSeq 6000 Integration v2.5.0 User Interaction, Validation and Troubleshooting.
NovaSeq 6000 Integration v2.5.0 has the following prerequisites:
Mount run data network-attached storage (NAS) share
Secret Util is installed
IPP is installed
NovaSeq 6000 Integration v2.5.0 supports both on-premise and cloud integrations. This integration is distributed as the following RPM packages:
BaseSpaceLIMS-novaseq-extensions
BaseSpaceLIMS-novaseq-sequencing-service (depends on BaseSpaceLIMS Automation Worker v1.x)
The BaseSpaceLIMS-novaseq-extensions RPM installs the following items:
Protocols and steps
Validation workflow
Database properties that configure the service
novaseq-extensions.jar file that provides sample sheet generation and other utility scripts
The BaseSpaceLIMS-novaseq-sequencing-service RPM installs the following items:
Bash scripts to run the novaseq_seqservice through systemctl
novaseq-sequencing.jar
novaseq-remote-extensions.jar file that contains scripts that need access to the run folders on the NAS
Use the following instructions to install the BaseSpaceLIMS-novaseq-extensions and BaseSpaceLIMS-novaseq-sequencing-service RPMs on the Clarity LIMS server.
This version of the integration only supports a single active search-replace pair of each type. The sampleSheetPathPrefixSearchReplaceSuffixes and netPathPrefixSearchReplaceSuffixes properties must contain a single number instead of a comma-separated list of numbers.
The sample sheet, run recipe, and run output folders can all be configured independently and do not have to be on the same mount or share. For examples of valid configuration mount points and properties, refer to Example Network Folder Structure and Database Properties.
The BaseSpaceLIMS-novaseq-extensions RPM must be installed on the Clarity LIMS server. The BaseSpaceLIMS-novaseq-sequencing-service RPM can be installed remotely on another server within the network.
This version of the integration only supports a single active search-replace pair of each type. The sampleSheetPathPrefixSearchReplaceSuffixes and netPathPrefixSearchReplaceSuffixes properties must contain a single number instead of a comma-separated list of numbers.
The sample sheet, run recipe, and run output folders can all be configured independently and do not have to be on the same mount or share. For examples of valid configuration mount points and properties, refer to Example Network Folder Structure and Database Properties.
For detailed descriptions of the steps and automations included in each protocol, and details on other components in the configuration, refer to NovaSeq 6000 Integration v2.5.0 Configuration.
For instructions on using the NovaSeq Validation Library Prep (NovaSeq 6000 v2.3) protocol, refer to NovaSeq 6000 Integration v2.5.0 User Interaction, Validation and Troubleshooting.
The NovaSeq instrument software includes the following components:
NovaSeq Control Software (NVCS) — Contains the user interface for setting up the sequencing run. Responsible for controlling the instrument and acquiring the images.
Real-Time Analysis v3 (RTA3) — Takes the images generated by the first module, processes, and analyzes them. Makes sure that data files are created and copied to the final destination folder.
Sequencing Analysis Viewer (SAV) — Displays the important quality metrics generated by the RTA3 software.
NovaSeq recipes — Provides system operation instructions for use with NovaSeq 6000 reagent kits for SP, S1, S2, and S4 flow cells.
Universal Copy Service — Copies output files to destinations such as final destination folder and/or BaseSpace Sequence Hub (when the instrument is configured for use with BaseSpace Sequence Hub).
For the NovaSeq API integration to work, the NovaSeq instrument must be able to communicate with Clarity LIMS through the API. Complete the following steps to configure the NVCS and confirm that you can access Clarity LIMS from the instrument.
Launch NVCS and wait for the initialization process to complete.
On the NVCS Settings page, do the following:
Under Mode Selection, select File-Based.
In the adjacent field, enter the network folder location that NVCS retrieves the JSON recipe from.
In the Output Folder field, enter the default output folder location.
Select Save to complete the configuration.
On the NVCS home page, you can now select Sequence to log into Clarity LIMS and start a run.
You must include the --enablerepo command line argument. Illumina Support provides the repo file and appropriate name to use.
For the on-premise installation, the Illumina Vault server is not available for public access.
If you are running the sequencing service and Automation Worker on the same instance, make sure that the instance is running a compatible version of Oracle Linux. For compatibility, refer to NovaSeq 6000 Integration v2.5.0 Release Notes.
You must include the --enablerepo command line argument. Illumina Support provides the repo file and appropriate name to use.
The configuration provided with this integration has been established to support NovaSeq 6000 lab processes. Any configuration changes to protocols or workflows, including renaming protocols, steps, and fields, could break processes.
Property
Description
novaseq.runSetupFolder
The directory where the Prepare Files for NovaSeq script writes run recipe files in the JSON format. This must be an existing folder and must be writable by the glsai user. For more information, refer to Prerequisites.
novaseq.sampleSheetPathPrefixSearch.SUFFIX
The directory where NVCS or analysis software looks for sample sheets. This directory is typically a Windows path or a network drive. The integration does not use this path directly, but the attachment filed in the run recipe file is determined by adding the sample sheet file name to this value.
novaseq.sampleSheetPathPrefixReplace.SUFFIX
The directory where the Prepare Files for NovaSeq script writes sample sheets. This must be an existing folder and must be writable by the glsai user. For more information, refer to Prerequisites.
novaseq.sampleSheetPathPrefixSearchReplaceSuffixes
This property must be set to the SUFFIX used in the novaseq.sampleSheetPathPrefixSearch.SUFFIX and novaseq.sampleSheetPathPrefixReplace.SUFFIX properties.
novaseq.seqservice.netPathPrefixSearch.SUFFIX
This property is the search prefix used to map the OutputRunFolder field in the RunParameters.xml file to the corresponding Linux directory. The OutputRunFolder is converted by replacing this string with the netPathPrefixReplace value. The value of this property is also in the output_folder field of the run recipe file.
novaseq.seqservice.netPathPrefixReplace.SUFFIX
The directory where the sequencing service looks for run folders. This must be an existing folder and must be writable by the glsai or glsjboss users, or by the claritylims group. For more information, refer to Prerequisites. The run folders must be in the configured directory. The service does not search sub-directories.
novaseq.seqservice.netPathPrefixSearchReplaceSuffixes
This property must be set to the single SUFFIX used in novaseq.seqservice.netPathPrefixSearch.SUFFIX and novaseq.seqservice.netPathPrefixReplace.SUFFIX.
Property
Description
novaseq.runSetupFolder
The directory where the Prepare Files for NovaSeq script writes run recipe files in the JSON format. This must be an existing folder and must be writable by the glsai user. For more information, refer to Prerequisites.
novaseq.sampleSheetPathPrefixSearch.SUFFIX
The directory where NVCS or analysis software looks for sample sheets. This directory is typically a Windows path or a network drive. The integration does not use this path directly, but the attachment filed in the run recipe file is determined by adding the sample sheet file name to this value.
novaseq.sampleSheetPathPrefixReplace.SUFFIX
The directory where the Prepare Files for NovaSeq script writes sample sheets. This must be an existing folder and must be writable by the glsai user. For more information, refer to Prerequisites.
novaseq.sampleSheetPathPrefixSearchReplaceSuffixes
This property must be set to the SUFFIX used in the novaseq.sampleSheetPathPrefixSearch.SUFFIX and novaseq.sampleSheetPathPrefixReplace.SUFFIX properties.
novaseq.seqservice.netPathPrefixSearch.SUFFIX
This property is the search prefix used to map the OutputRunFolder field in the RunParameters.xml file to the corresponding Linux directory. The OutputRunFolder is converted by replacing this string with the netPathPrefixReplace value. The value of this property is also in the output_folder field of the run recipe file.
novaseq.seqservice.netPathPrefixReplace.SUFFIX
The directory where the sequencing service looks for run folders. This must be an existing folder and must be writable by the glsai or glsjboss users, or by the claritylims group. For more information, refer to Prerequisites. The run folders must be in the configured directory. The service does not search sub-directories.
novaseq.seqservice.netPathPrefixSearchReplaceSuffixes
This property must be set to the single SUFFIX used in novaseq.seqservice.netPathPrefixSearch.SUFFIX and novaseq.seqservice.netPathPrefixReplace.SUFFIX.