Step 4: Load to Library Tube Strip (NovaSeq X Series Sequencing v1.0)

1. 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.
2. Add the pools to the Ice Bucket and select View Ice Bucket.
3. On the Ice Bucket screen, select Begin Work to trigger the Validate Analysis Configurations and Register Step Started automation.

This 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.

The automation also registers the start time of the step by publishing messages to CLPA through the ICA LINK service. The portion of the script used for start time registration is used only for CLPA support.

4. On the Placement screen, do as follows.
a. Drag the pool into the Library 8-tube Strip in the Placed Samples area on the right.
b. Scan or type the barcode of the Library 8-tube Strip into the Library 8-tube Strip container name field.
c. 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.

5. 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

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

¹ The Local run mode generates the sample sheet. The Run/Analysis configuration is imported manually to the NovaSeq X Series instrument through the sample sheet. The sample sheet 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 Cloud run mode sends the Run/Analysis configuration to ICA. This configuration is downloaded to the NovaSeq X Series instrument through ICA to start the run. The downstream secondary analysis is done in 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.

6. 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.
• Creates the planned run and sends it to ICA when the Run Mode is Cloud.
• Generates the sample sheet and attaches it to the placeholder in the Files area on the Record Details screen for all Run Mode types.
7. 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.

8. 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 Configuration and User Interaction.
9. 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.