IGA NovaSeq Sequencing v2.1
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Last updated
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IGA NovaSeq Sequencing v2.1 contains preset protocols that support the setup of previously prepared libraries used with the NovaSeq instrument. These protocols can be used with the IGA Library Prep Automated v2.1 workflow. They also provide the following functions:
File-based integration with the Hamilton robots that are used for liquid handling steps.
Automated calculation of sample and buffer volumes.
Automated calculation or display of reagents.
When required, automatic step transition.
Protocol Type = Library Prep
Next Steps Configuration
Master Step Name = Define Run Format (IGA v2.1)
Step Type = No Outputs
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table
Column Headers
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
Well
Built-in
Derived Sample
Reagent Name
Built-in
Derived Sample
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Expanded View Fields
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
LIMS ID (Container)
Built-in
Project
Project Name
Built-in
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table
Column Headers
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
Well
Built-in
Derived Sample
Reagent Name
Built-in
Derived Sample
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Expanded View Fields
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
LIMS ID (Container)
Built-in
Project
Project Name
Built-in
Step Data (Master Step Fields)
Field Name
Field Type
Options
Additional Options and Dropdown Items
IGA WF Build Version
Text
Read Only
Default = 2.1.0.1
Quick reference checklist: Define Run Format.
Multiline Text
Read Only
Defaults = User tasks for this page: - Below, assign 'Loading Workflow Type', 'Flowcell Type', and the NovaSeq 'Final Loading Concentration (pM)'. - Click 'Next Steps'.
Step File Placeholders
Log file - Automatically attached
Sample Table (Table Columns - Global Fields)
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
LIMS ID (Container)
Built-in
Container
Well
Built-in
Derived Sample
Adjusted Per Sample Volume (ul)
Numeric
Read Only
Decimal Places Displayed = 2
Derived Sample
Final Loading Concentration (pM)
Numeric Dropdown
Required Field
Custom Entries
Presets
225
400
Decimal Places Displayed = 0
Derived Sample
Flowcell Type
Text Dropdown
Required Field
Presets
SP
S1
S2
S4
Derived Sample
Loading Workflow Type
Text Dropdown
Required Field
Presets
NovaSeq Standard
NovaSeq Xp
[Remove from workflow]
Derived Sample
Minimum Molarity (nM)
Numeric
Decimal Places Displayed = 2
Derived Sample
Normalized Molarity (nM)
Numeric
Decimal Places Displayed = 2
Derived Sample
Per Sample Volume (ul)
Numeric
Read Only
Decimal Places Displayed = 2
Derived Sample
Sample Name
Built-in
Derived Sample
Warning
Text
Read Only
Project
Project Name
Built-in
Protocol Type = Library Prep
Next Steps Configuration
Master Step Name = Make Bulk Pool for NovaSeq Standard (IGA v2.1)
Step Type = Pooling
Aliquot Generation = Fixed, 1
Naming Convention = {PoolName}
Control Types
PhiX v3
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table
Column Headers
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
Well
Built-in
Derived Sample
Flowcell Type
Text Dropdown
Required Field
Presets
SP
S1
S2
S4
Derived Sample
Normalized Molarity (nM)
Numeric
Decimal Places Displayed = 2
Derived Sample
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Expanded View Fields
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
LIMS ID (Container)
Built-in
Project
Project Name
Built-in
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table
Column Headers
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
Well
Built-in
Derived Sample
Flowcell Type
Text Dropdown
Required Field
Presets
SP
S1
S2
S4
Derived Sample
Normalized Molarity (nM)
Numeric
Decimal Places Displayed = 2
Derived Sample
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Expanded View Fields
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
LIMS ID (Container)
Built-in
Project
Project Name
Built-in
Label Uniqueness = On
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Defaults
Well Sort Order = Column
Placement Pattern = Column
Destination Containers
96 well plate
Step Data (Master Step Fields)
Field Name
Field Type
Options
Additional Options and Dropdown Items
Number of Flowcells to Sequence
Numeric
Required Field
Default = 1
Range From 1 to 10
Decimal Places Displayed = 0
Minimum Per Sample Volume (ul)
Numeric
Required Field
Default = 2
Decimal Places Displayed = 2
Liquid Handling Instrument ID
Text
Read Only
Apply Correction Factors
Text Dropdown
Required Field
Presets
None
IDPF
Custom
Default = None
Quick reference checklist: Make Bulk Pool for NovaSeq Standard. i) pre Liquid Handler
Multiline Text
Read Only
Defaults = User tasks for this page: - Select 'Number Of Flowcells' and 'Min Per Sample Volume (ul)' - 'Run' the Make Bulk Pool automation, and download resulting Hamilton Make Bulk Pool Input File
Quick reference checklist: Make Bulk Pool for NovaSeq Standard. ii) post Liquid Handler
Multiline Text
Read Only
Defaults = User tasks for this page: - After running MakeBPP on the liquid handler, upload resulting Hamilton Output File - Click on 'Next Steps'
Step File Placeholders
Calculation Log - Automatically attached
Hamilton Make Bulk Pool Input File - Automatically attached
Hamilton file log - Automatically attached
Hamilton Output File - Manually uploaded
Custom Correction Factors - Manually uploaded
Index Correction Log - Automatically attached
Sample Table (Table Columns - Global Fields)
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
LIMS ID (Container)
Built-in
Container
Well
Built-in
Derived Sample
Flowcell Type
Text Dropdown
Required Field
Presets
SP
S1
S2
S4
Derived Sample
Loading Workflow Type
Text Dropdown
Required Field
Presets
NovaSeq Standard
NovaSeq Xp
[Remove from workflow]
Derived Sample
NaOH Volume (ul)
Numeric
Read Only
Decimal Places Displayed = 2
Derived Sample
RSB Volume (ul)
Numeric
Read Only
Decimal Places Displayed = 2
Derived Sample
Sample Name
Built-in
Derived Sample
Tris-HCl Volume (ul)
Numeric
Read Only
Decimal Places Displayed = 2
Derived Sample
Volume of Pool to Denature (ul)
Numeric
Read Only
Decimal Places Displayed = 0
Project
Project Name
Built-in
Master Step Name = Dilute and Denature (IGA v2.1)
Step Type = Standard
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}
Reagent Kits
NaOH
Supplier = Illumina
Resuspension Buffer (RSB)
Supplier = Illumina
Tris-HCl
Supplier = Illumina
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table
Column Headers
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
Well
Built-in
Derived Sample
Flowcell Type
Text Dropdown
Required Field
Presets
SP
S1
S2
S4
Derived Sample
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Expanded View Fields
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container