Considerations when using DRAGEN TruSight Oncology 500 Solid v2.1 Sample Sheet

Considerations when using DRAGEN TruSight Oncology 500 (TSO500) Solid v2.1 Sample Sheet

1. Where can we download the template ? In the resource bundle installed with the software (/staging/illumina/DRAGEN_ TSO500/resources/sampleSheet/)

2. Considerations when using DRAGEN TruSight Oncology 500 Solid v2.1 Sample Sheet

• DRAGEN TSO500 solid v2.1 requires a v2 sample sheet; v1 sample sheets are not supported.

• Default name: SampleSheet.csv

• Default location: top level of the run folder or FASTQ folder

• Sample IDs must be unique and meet the following criteria:

  • 1 to 40 characters in length, no spaces

  • Alphanumeric, dash, and underscore characters only

  • An underscore or dash must have an alphanumeric character immediately before and after

  • Cannot be called all, default, none, unknown, undetermined, stats, or reports.

  • It is recommended that the sample ID be based on the pair ID. Example: -DNA,- RNA.

• A “Lane” column in the [BCLConvert_Data] section is required only if user is using the XP flow cell and an index pair is used for more than one sample, the samples must be distinguished by lane separation. Use the lane column to specify sample location per lane. User must specify a lane for each sample. If a sample is in multiple lanes, user must use multiple rows for each sample (one row per lane).

• If the “Lane” column will not be used, do not include the column in the sample sheet.

• For HRD samples, see note below about the new “Sample_Feature” column.

• Make sure the sample sheet does not contain any blank lines at the bottom. The lines will cause the analysis to exit unsuccessfully.

*\\\Notes

v2.1 update: a new column has been added to the [TSO500S_Data] section of the sample sheet: Sample_Feature

• The Sample_Feature column is only required if analyzing HRD-enriched samples.

• For a HRD sample:

  • Sample_Type = DNA

  • Sample_Feature = HRD

• Only DNA sample types can have the sample feature HRD.

• The inclusion of the Sample_Feature column prompts the software to check for a valid TSO500_HRD license.

v2.1 update: removed support for v1 sample sheets.

If using the NovaSeq v1.5 chemistry, the i5 indexes need to be reverse complemented in the sample sheet. If the user forgets to do this, or reverse complements in the sample sheet but used v1 chemistry, the software will show an error early on, because it will check which chemistry was used and make sure the corresponding index orientation was used.

The v2 sample sheet allows for all libraries to be included in FASTQ generation, while including only selected libraries in the subsequent DRAGEN TSO500 analysis.

• To include samples in the FASTQ generation (BCLConvert) step, list them in the [BCLConvert_Data] section of the sample sheet.

• To include samples in the DRAGEN TSO500 analysis step, list them in the [TSO500S_Data] section of the sample sheet.

• When starting DRAGEN TSO 500 analysis from BCLs, TSO 500 samples need to be listed in both the [BCLConvert_Data] and [TSO500S_Data] sections of the sample sheet.

Index 2 (i5) sequence orientation:

• For NovaSeq 6000 v1.5 chemistry: Enter sequences in the reverse complement orientation

• NextSeq 500/550: Enter sequences in the reverse complement orientation

More information regarding Sample Sheet requirements for DRAGEN TSO500 Software v2.1 can be found in DRAGEN TSO500 Analysis Software v2.1 User Guide ( page 7,8)

For any feedback or questions regarding this article (Illumina Knowledge Article #7419), contact Illumina Technical Support [e-mail].