Protocol Type = Sample Prep
Next Steps Configuration
Master Step Name = Hybridize Probes for Stranded Total RNA Prep v1.0
Step Type = Standard
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Deplete rRNA v1.0
Step Type = No Outputs
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Remove Probes v1.0
Step Type = No Outputs
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Clean Up RNA v1.0
Step Type = Standard
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Fragment and Denature RNA v1.0
Step Type = No Outputs
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Row
Table Columns - Global Fields
Master Step Name = Synthesize First Strand cDNA v1.0
Step Type = No Outputs
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Synthesize Second Strand cDNA v1.0
Step Type = Standard
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}
Reagent Kits
Agencourt AMPure XP, 5 ml
Supplier = Beckman Coulter
Catalog Number = A63880
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Protocol Type = Library Prep
Next Steps Configuration
Master Step Name = Adenylate 3ʹ Ends - RNA Prep v1.0
Step Type = No Outputs
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Ligate Anchors - RNA Prep v1.0
Step Type = No Outputs
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Clean Up Fragments - RNA Prep v1.0
Step Type = Standard
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}
Reagent Kits
Agencourt AMPure XP, 5 ml
Supplier = Beckman Coulter
Catalog Number = A63880
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Amplify Library - RNA Prep v1.0
Step Type = Add Labels
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}-{AppliedReagentLabels}
Reagent Kits
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Label Groups
IDT for Illumina RNA UD Indexes Set A Ligation
IDT for Illumina RNA UD Indexes Set A-D Ligation
IDT for Illumina RNA UD Indexes Set B Ligation
IDT for Illumina RNA UD Indexes Set C Ligation
IDT for Illumina RNA UD Indexes Set D Ligation
Illumina RNA UD Indexes Set A B C D Ligation
Illumina RNA UD Indexes Set A Ligation
Illumina RNA UD Indexes Set B Ligation
Illumina RNA UD Indexes Set C Ligation
Illumina RNA UD Indexes Set D Ligation
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Master Step Name = Clean Library v1.0
Step Type = Standard
Derived Sample Generation = Fixed, 1
Naming Convention = {InputItemName}
Reagent Kits
Agencourt AMPure XP, 5 ml
Supplier = Beckman Coulter
Catalog Number = A63880
Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus
Supplier = Illumina
Website = www.illumina.com
Defaults
Sample Grouping = Group by Containers
Well Sort Order = Column
Sample Table (Column Headers)
Step Data (Master Step Fields)
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
DB1 (uL)
Numeric
Decimal Places Displayed = 0
DP1 (uL)
Numeric
Decimal Places Displayed = 0
Instruction Notes
Multiline Text
Read Only
Default = Add 4 µl Hybridize Probe Master Mix to each well.
Thermal Cycler Program
Text
Default = HYB_DP1
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheat lid option and set to 100°C Reaction volume is 15 µl u 95° for 2 minutes Decrease 0.1°C per second until the temperature reaches 37°C Hold at 37°C
Total Samples
Numeric
Default = 0
Decimal Places Displayed = 0
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
Instruction Notes
Multiline Text
Read Only
Default = Add 5 µl rRNA Depletion Master Mix to each well.
RDB (uL)
Numeric
Decimal Places Displayed = 0
RDE (uL)
Numeric
Decimal Places Displayed = 0
Thermal Cycler Program
Text
Default = RNA_DEP
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = RNA_DEP program on the thermal cycler:Choose the preheat lid option and set to 100°C Reaction volume is 20 µl 37°C for 15 minutes Hold at 4°C
Total Samples
Numeric
Default = 0
Decimal Places Displayed = 0
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
Instruction Notes
Multiline Text
Read Only
Default = Add 10 µl Probe Removal Master Mix to each well.
PRB (uL)
Numeric
Decimal Places Displayed = 0
PRE (uL)
Numeric
Decimal Places Displayed = 0
Thermal Cycler Program
Text
Default = PRB_REM
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = PRB_REM program on the thermal cycler: Choose the preheat lid option and set to 100°C Reaction volume is 30 µl 37°C for 15 minutes 70°C for 15 minutes Hold at 4°C
Total Samples
Numeric
Default = 0
Decimal Places Displayed = 0
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
Instruction Notes
Multiline Text
Read Only
Default = Add 8.5 µl EPH3 to each well.
Thermal Cycler Program
Text
Default = DEN_RNA
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheat lid option and set to 100°C Reaction volume is 17 µl 94°C for 2 minutes Hold at 4°C
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
FSA (uL)
Numeric
Decimal Places Displayed = 0
Instruction Notes
Multiline Text
Read Only
Default = Add 8 uL of First Strand Synthesis Master Mix to each sample.
RVT (uL)
Numeric
Decimal Places Displayed = 0
Thermal Cycler Program
Text
Default = FSS
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheat lid option and set to 100°C Reaction volume is 25 µl 25°C for 10 minutes 42°C for 15 minutes 70°C for 15 minutes Hold at 4°C
Total Samples
Numeric
Default = 0
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
80% EtOH Prep Date
Date
Thermal Cycler Program
Text
Default = SSS
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheat lid option and set to 40°C Reaction volume is 50 µl u 16°C for 1 hour Hold at 4°C
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
Instruction Notes
Multiline Text
Read Only
Default = Add 12.5 µl ATL4 to each well.
Thermal Cycler Program
Text
Default = ATAIL
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheat lid option and set to 100°C Reaction volume is 30 µl 37°C for 30 minutes 70°C for 5 minutes Hold at 4°C
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
Add Anchors Instruction Notes
Multiline Text
Default = Volume for Sample Input <= 100 ng (µl) 1. Add 2.5 (µl) RSB 2. Add 2.5 (µl) RNA Index Anchors 3. Add 2.5 (µl) LIGX Volume for Sample Input > 100 ng (µl) 1. Add 0 (µl) RSB 2. Add 5 (µl) RNA Index Anchors 3. Add 2.5 (µl) LIGX
Instruction Notes
Multiline Text
Read Only
Default = Add 5 µl STL to each well.
Thermal Cycler Program
Text
Default = LIG
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheated lid option and set to 100°C Reaction volume is 38 µl 30°C for 10 minutes Hold at 4°C
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
80% EtOH Prep Date
Date
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
PCR Cycles
Text Dropdown
Custom Entries
Presets
15
13
10
Default = 15
Recommendations for PCR Cycle Number
Multiline Text
Read Only
Default = Use 15 cycles for 10 ng of input DNA Use 13 cycles for 100 ng of input DNA Use 10 cycles for 1000 ng of input DNA
Thermal Cycler Program
Text
Default = PCR
Thermal Cycler Program Notes
Multiline Text
Read Only
Default = Choose the preheat lid option and set to 100°C Reaction volume is 50 µl u 98°C for 30 seconds X cycles of: 98°C for 10 seconds 60°C for 30 seconds 72°C for 30 seconds 72°C for 5 minutes Hold at 4°C for ≤ 16 hours
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
Sample Name
Built-in
Project
Project Name
Built-in
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
Sample Name
Built-in
Derived Sample
Waiting
Built-in
Project
Project Name
Built-in
Field Name
Field Type
Options
Additional Options and Dropdown Items
80% EtOH Prep Date
Date
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
Sample Name
Built-in
Project
Project Name
Built-in
