Sample Sheet Creation in BaseSpace Run Planning tool

How to Create TSO 500 Sample Sheets in BaseSpace Run Planning tool

The BaseSpace Sequence Hub Run Planning tool is available, and is used to generate a valid sample sheet in v2 format for use on a TSO 500 supported sequencer for both ICA and Standalone DRAGEN Server analysis options. Filling out the form on the user interface will produce a exportable sample sheet with the required fields filled in. Refer to ICA Auto-launch Sample Sheet Requirements for descriptions of fields that appear in ICA sample sheets.

The sections below represent each step in the BaseSpace Run Planning tool.

Note that NovaSeq X Series has a different run set up configuration screen than other instrument platforms. TSO 500 does not support multi analysis, and in order to run TSO 500 on NovaSeq X Series, enter the appropriate Read 1, Read 2, Index 1 and Index 2 described in the instructions below.

BaseSpace Run Planning tool cannot generate a valid sample sheet for NovaSeq 6000Dx Analysis Application. Refer to Sample Sheet Requirements page to create a valid sample sheet.

Step 1: Run Settings

Parameter Name
Required
Description

Run Name

Required

Run Name can contain 255 alphanumeric characters, dashes, underscores, periods, and spaces; and must start with an alphanumeric, a dash or an underscore.

Run Description

Optional

Run Description can contain 255 characters except square brackets, asterisks, and commas.

Instrument Platform

Required

Choose from TSO 500 supported instruments:

  • NextSeq 500/550

  • NextSeq 1000/2000

  • NovaSeq 6000/6000Dx

  • NovaSeq X Series

Secondary Analysis

Required

  • BaseSpace/Illumina Connected Analytics (to generate sample sheet for cloud analysis)

  • Local

Read 1

Required on Instrument Platform NovaSeq X Series

  • Fill with value 101 for TSO 500 analysis

Index 1

Required on Instrument Platform NovaSeq X Series

  • Fill the value depending on the TSO 500 assay used:

  • TSO 500 HT: 10

  • TSO 500: 8

Index 2

Required on Instrument Platform NovaSeq X Series

  • Fill the value depending on the TSO 500 assay used:

  • TSO 500 HT: 10

  • TSO 500: 8

Read 2

Required on Instrument Platform NovaSeq X Series

  • Fill with value 101 for TSO 500 analysis

Sample Container ID

Optional

  • Unique Identifier for the container that holds the sample

Step 2: Configuration

Note: On NovaSeq X Series, this page is called "Configuration 1". The right hand corner of the UI displays the Read 1, Read 2, Index 1 and Index 2 entered on the previous run settings screen.

Parameter Name
Required
Description

Application*

Required

  • DRAGEN TruSight Oncology 500 Analysis Software - 2.6.0 (with HRD)

  • DRAGEN TruSight Oncology 500 Analysis Software - 2.6.0

Description

Optional

Optional text field

Library Prep Kit

Required

  • TruSight Oncology 500

  • TruSight Oncology 500 High Throughput

Index Adapter Kit

Required

TSO 500:

  • TruSight Oncology 500 (NovaSeq6000Dx with S1, S2, S4, or SP Flow Cell, NextSeq1000/2000, NovaSeqX Series)

  • TruSight Oncology 500 (NovaSeq6000, NextSeq)

TSO 500 HT:

  • TruSight Oncology 500 High Throughput (NovaSeq6000Dx with S1, S2, S4, or SP Flow Cell, NextSeq1000/2000, NovaSeqX Series)

  • TruSight Oncology 500 High Throughput (NovaSeq6000, NextSeq)

Step 3: Sample Settings

Users can manually enter sample information, or download a template file to bulk upload sample information. Users can import the completed template or a compatible sample sheet.

Parameter Name
Required
Description

Read Lengths: Read 1 and Read 2

Required Not applicable on NovaSeq X Series

Auto filled with the standard values, but can be optionally overwritten.

Override Cycles

Required on NovaSeq X Series

Entered based on Run Settings read lengths & index 1 / index 2

Lane Usage

Not applicable on NovaSeq X Series or NextSeq 1000 / 2000

Checkbox allows users to apply the same lane across samples.

Lane

Required if Lane Usage is unchecked Not applicable on NextSeq 1000 / 2000

Specify lanes for each sample. The unmarked checkbox at the top of the dropdown selects all lanes.

Pair ID

Required

The identifier used to pair DNA and RNA samples in a run. The field is mandatory whether a sample is part of a pair, or not.

To note: The Sample ID field in the generated samplesheet will be auto-filled based on the Pair ID values captured. “_dna” and “_rna” (for DNA and RNA samples respectively) will be appended to the Pair ID value to create the Sample ID.

DNA Index ID

Required

Index set ID options are based on selected Index Adapter Kit

DNA Sample Feature

Required for TSO 500 HRD

Column appears when TSO 500 HRD application is selected. Enter for HRD enriched DNA Samples

RNA Index ID

Required

Index set ID options are based on selected Index Adapter Kit

Project

Optional

Optional field to describe the associated project

Starts from Fastq

Required

True or False

If auto-launching TSO 500 from BCL files, set the value to False. If auto-launching TSO 500 from FASTQ after auto-launching BCL Convert, set the value to True.

DNA Barcode Mismatches Index 1**

DNA Barcode Mismatches Index 2**

RNA Barcode Mismatches Index 1**

RNA Barcode Mismatches Index 2**

Required on NovaSeq X

Default value is set to 1.

These fields are required by NovaSeq X and represent BCL Convert settings for index diversity checks when demultiplexing. These values are not used in TSO 500 analysis.

Step 4: Run Review

Once all details are captured and pass validation, the user can review the details on the Run Review screen. From here they can choose to edit details in previous screens or export the sample sheet. Once completed, press the Cancel button to finish run planning.

Note: once leaving this screen, the run and sample sheet will not be accessible.

For NovaSeqX Plus users, the run can be saved as a draft or as a planned run (via “Save as Draft” and “Save as Planned” buttons respectively). Either selection will save the run to the Planned Runs screen on BaseSpace. There is no option to export the sample sheet on this screen.

Planned Runs Screen (NovaSeq X Series only)

The Planned Runs screen lists all planned or drafted runs. Users can set drafted runs to planned, export the sample sheet, and edit or delete a run on this screen.

Once the run is saved as Planned, it will appear on the NovaSeq X Series instrument where it can be selected for sequencing.

For more information on run planning, refer to the BaseSpace Sequence Hub support site page.

Guided Examples

Please review these guided examples of analysis workflows that include a step of setting up a run in BaseSpace Run Planning tool:

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