# Sample Sheet Requirements

The DRAGEN Single Cell RNA software has optional and required fields in addition to general sample sheet requirements. Below is a description of the fields in each section.

{% hint style="info" %}
The preferred method for creating sample sheets is to use [Run Planning in BaseSpace Sequence Hub](https://help.multiomics.illumina.com/dragen-single-cell-rna/run-set-up-in-bssh/run-planning)
{% endhint %}

## \[Sequencing\_Settings]

<table><thead><tr><th width="186">Parameter</th><th width="132">Required?</th><th>Details</th></tr></thead><tbody><tr><td>LibraryPrepKits</td><td>Required</td><td>Accepted values are: IlluminaSingleCell3RNAPrep</td></tr></tbody></table>

## \[BCLConvert\_Settings]

<table><thead><tr><th width="195">Parameter</th><th width="129">Required?</th><th>Details</th></tr></thead><tbody><tr><td>SoftwareVersion</td><td>Required</td><td>The DRAGEN component software version. DRAGEN Single Cell RNA software requires 4.4.0.</td></tr><tr><td>NoLaneSplitting</td><td>Required</td><td>TRUE for DRAGEN Single Cell RNA software</td></tr><tr><td>TrimUMI</td><td>Required</td><td>0 for DRAGEN Single Cell RNA software</td></tr><tr><td>OverrideCycles</td><td>Required</td><td>U45;I10;I10;Y72 for DRAGEN Single Cell RNA software. May be different if running multiple applications in a single run.</td></tr><tr><td>FastqCompressionFormat</td><td>Required</td><td>gzip</td></tr></tbody></table>

## \[BCLConvert\_Data]

<table><thead><tr><th width="220">Parameter</th><th width="151">Required?</th><th>Details</th></tr></thead><tbody><tr><td>Sample_ID</td><td>Required</td><td>Must match a Sample_ID listed in the [Cloud_DragenSingleCellRna_Data] and [Cloud_Data] section.</td></tr><tr><td>Index</td><td>Required</td><td>Index 1 sequence</td></tr><tr><td>Index2</td><td>Required</td><td>Index 2 sequence</td></tr><tr><td>Lane</td><td>Only for NovaSeq 6000/6000 Dx workflow</td><td>Indicates which lane corresponds to a given sample. Enter a single numeric value per row. Cannot be empty, i.e. the analysis fails if the Lane column is present without a value in each row.</td></tr></tbody></table>

## \[Cloud\_DragenSingleCellRna\_Settings]

<table><thead><tr><th width="207">Parameter</th><th width="137">Required?</th><th>Details</th></tr></thead><tbody><tr><td>SoftwareVersion</td><td>Required</td><td>The DRAGEN component software version. DRAGEN Single Cell RNA software requires 4.4.0.</td></tr><tr><td>EnablePipseqMode</td><td>Required</td><td>TRUE for Illumina Single Cell 3’ RNA kit. Maps to --scrna-enable-pipseq-mode in command line arguments.</td></tr><tr><td>ReferenceGenomeDir</td><td>Required</td><td>Location of reference genome TAR containing a DRAGEN hash table and optionally a GTF.</td></tr><tr><td>BarcodeRead</td><td>Required</td><td>Read1 for Illumina Single Cell 3’ RNA kit</td></tr><tr><td>RnaLibraryType</td><td>Required</td><td>SF for Illumina Single Cell 3’ RNA kit (stranded forward). Maps to --rna-library-type in command line arguments.</td></tr><tr><td>BarcodePosition</td><td>Required</td><td>0_7+11_16+20_25+31_38 for Illumina Single Cell 3’ RNA kit. Maps to --scrna-barcode-position in command line arguments.</td></tr><tr><td>UmiPosition</td><td>Required</td><td>39_41 for Illumina Single Cell 3’ RNA kit. Maps to --scrna-umi-position in command line arguments.</td></tr></tbody></table>

## \[Cloud\_DragenSingleCellRna\_Data]

<table><thead><tr><th width="213">Parameter</th><th width="135">Required?</th><th>Details</th></tr></thead><tbody><tr><td>Sample_ID</td><td>Required</td><td>Must match a Sample_ID listed in the [BCLConvert_Data] and [Cloud_Data] section.</td></tr></tbody></table>

## \[Cloud\_Settings] for Auto-launch

<table><thead><tr><th width="225">Parameter</th><th width="128">Required?</th><th>Details</th></tr></thead><tbody><tr><td>GeneratedVersion</td><td>Not Required</td><td>The cloud version used to create the sample sheet. Optional if manually updating a sample sheet. (ex: 1.17.0.202411192008).</td></tr><tr><td>Cloud_Workflow</td><td>Not Required</td><td>ica_workflow_1</td></tr><tr><td>BCLConvert_Pipeline</td><td>Required</td><td><p>The value is a universal record number (URN). The valid value is:</p><p>urn:ilmn:ica:pipeline:730df76f-715a-45bf-9500-e6e0ce1ab224#BclConvert_v4_3_13</p></td></tr><tr><td>Cloud_DragenSingleCellRna_Pipeline</td><td>Required</td><td><p>The value is a URN in the following format:</p><p>urn:ilmn:ica:pipeline:b3c5ab5f-2853-4873-93c4-61a807f844a7#DRAGEN_Single_Cell_RNA_4-4-2_-_Sequencer_Integration_Only</p></td></tr></tbody></table>

## \[Cloud\_Data] for Auto-Launch

<table><thead><tr><th width="229">Parameter</th><th width="128">Required?</th><th>Details</th></tr></thead><tbody><tr><td>Sample_ID</td><td>Required</td><td>Must match a Sample_ID listed in the [BCLConvert_Data] and [Cloud_DragenSingleCellRna_Data] section.</td></tr><tr><td>ProjectName</td><td>Not Required</td><td>The BaseSpace Sequence Hub project name</td></tr><tr><td>LibraryName</td><td>Not Required</td><td>Combination of sample ID and index values in the following format: sampleID_Index_Index2.</td></tr><tr><td>LibraryPrepKit</td><td>Required</td><td>The Library Prep Kit used</td></tr><tr><td>IndexAdapterKitName</td><td>Required</td><td>The Index Adapter Kit used</td></tr></tbody></table>