Create one sample sheet for each flow cell. There are two ways to generate a sample sheet:
• Use the Singular Genomics online Sample Sheet Generator (recommended).
• Generate a sample sheet manually as described below.
NOTE
You do not have to upload a sample sheet. You can also define the samples manually when you set up the run on the ICS, or not provide sample data at all. However, without index information the Instrument does not perform on-instrument demultiplexing.
1. Download a sample sheet template from the Instrument, C:\Program Files (x86)\Singular Genomics\G4 Instrument Control Software\RunTemplates.
NOTE
If you do not plan on specifying demultiplexing options, use the template without Demux section (FlowCellTemplate.csv). Do not use FlowCellTemplateDemuxFields.csv, since you need to edit the demultiplexing options in order for that template to work properly.
2. Fill out the following information for the Header section. Fields marked with an asterisk * are required.
Field
Description
Date
Date of sample sheet edit.*
User Name
Name of operator.
User Email
Email of operator.
Workflow
Allows you to track the analysis pipeline.
Assay
Allows you to track the library prep used.
Run Notes
Any other notes for the run.
3. Fill out the following information for the Settings section. Fields marked with an asterisk * are required.
Note
Do not enter an index length that is longer than the physical length of the indices. This may interfere with the binding of the sequencing primers in subsequent sequencing steps.
Description
[read 1]
Length of read 1 in bp, maximum 150 bp.*
[read 2]
Length of read 2 in bp, maximum 150 bp.
[Index 1]
Length of Index 1 in bp, minimum 8 bp, maximum 24 bp.
[Index 2]
Length of Index 2 in bp, minimum 8 bp, maximum 24 bp.
Custom Primer Read1 (Y)
If a custom sequencing primer is needed for read 1 enter Y. If not, leave blank.
Custom Primer Read2 (Y)
If a custom sequencing primer is needed for read 2 enter Y. If not, leave blank.
Custom Primer Index1 (Y)
If a custom sequencing primer is needed for Index 1 enter Y. If not, leave blank.
Custom Primer Index2 (Y)
If a custom sequencing primer is needed for Index 2 enter Y. If not, leave blank.
Note
Custom sequencing primers can only be set at the run level, you cannot use custom sequencing primers on one flow cell and Singular Genomics sequencing primers on another flow cell within the same run. Note also that different lanes within a flow cell cannot have different sequencing primers.
4. Fill out the following information for the samples section. Note that within a single lane, no two samples can have the same combination of Sample_ID, Index1_Name, Index2_Name, Index1_Sequence and Index2_Sequence. Fields marked with an asterisk * are required.
Description
Sample_ID
Sample ID.*
Index1_Name
Name of Index 1. Leave empty if no Index 1 is used.
Index1_Sequence
Sequence of Index 1. Required for demultiplexing. Leave empty if no Index 1 is used.
Index2_Name
Name of Index 2. Leave empty if no Index 2 is used
Index2_Sequence
Sequence of Index 2. Required for demultiplexing. Leave empty if no Index 2 is used
Lane
Lane number, a single number from 1 to 4. Required.
Lane_Name
Lane name.
Project
Allows you to organize libraries in a project.
Loading_Concentration
Loading concentration on lane.
Application
Allows you to track the library prep used.
Notes
Any other notes for the sample library.
Reference
Library reference.
Note
If you want to sequence a library in multiple lanes, copy the line for that library in the sample sheet and change the lane number.
5. Name the sample sheet and save as CSV file (*.csv) in the SampleData folder or in a location available on the same network as the instrument. If you save the sample sheet on a network location, you may need to specify the permissions required for the folder on the instrument.
Note
For custom demultiplexing options, add a Demux section in between the Settings and Data sections and specify options as described here: How Do I Specify Custom Demultiplexing in the Sample Sheet?.