Illumina RNA Prep with Enrichment (L) Tagmentation v1.1
Overview
The Illumina RNA Prep with Enrichment (L) Tagmentation workflow includes the following functionality.
Preset Illumina RNA Prep with Enrichment (L) Tagmentation protocols that support the generation of enriched libraries for dual-indexed, paired-end sequencing.
Automated calculation of sample and buffer volumes.
Automated calculation or display of reagents at every step in the protocol.
Automatic step transition when required.
Automatic placement of samples when necessary.
Automated assignment of QC Pass/Fail, based on user-selected threshold values.
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
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Naming Convention = {InputItemName}
Reagent Kits
Agencourt AMPure XP, 5 ml
Supplier = Beckman Coulter
Catalog Number = A63880
Illumina RNA Prep with Enrichment (L) Tagmentation
Supplier = Illumina
Catalog Number = 16 - 20040536, 96 - 20040537
Sample Table (Column Headers)
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
LIMS ID (Container)
Built-in
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
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Illumina RNA Prep with Enrichment (L) Tagmentation
IDT for Illumina DNA-RNA UD Indexes Set C Tagmentation
IDT for Illumina DNA-RNA UD Indexes Set D Tagmentation
Illumina DNA-RNA UD Indexes Set A Tagmentation
Illumina DNA-RNA UD Indexes Set B Tagmentation
Illumina DNA-RNA UD Indexes Set C Tagmentation
Illumina DNA-RNA UD Indexes Set D Tagmentation
Default = Add 40 µl PCR Master Mix to each well.
Nuclease-free ultrapure water (uL)
Numeric
Decimal Places Displayed = 0
PCR Cycles
Text Dropdown
Custom Entries
Presets
14 cycles for High-quality RNA with DV200 > 80%
17 cycles for FFPE and RNA with DV200 < 80%
Thermal Cycler Program
Text
Default = TAG_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 72°C for 3 minutes u 98°C for 3 minutes
X cycles of: u 98°C for 20 seconds u 60°C for 30 seconds
72°C for 1 minute u 72°C for 3 minutes
Hold at 10°C
Total Samples
Numeric
Default = 0
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Naming Convention = {InputItemName}
Reagent Kits
Agencourt AMPure XP, 5 ml
Supplier = Beckman Coulter
Catalog Number = A63880
Illumina RNA Prep with Enrichment (L) Tagmentation
Bioanalyzer XML Result File (required) - Manually uploaded
Result File (optional) - Manually uploaded
PDF Summary File (optional) - Manually uploaded
Bioanalyzer XML Parsing Log File - Automatically attached
QC Assignment Log File - Automatically attached
QC Assignment Report - Automatically attached
Sample Table
Enable QC Flags = Yes
Sample Display Default = Expand
Well Sort Order = Column
File Column Options
File Column Display = Show
File Attachment Method = Manual
Table Columns - Global Fields
Naming Convention = {InputItemName}
Reagent Kits
Illumina RNA Prep with Enrichment (L) Tagmentation
Supplier = Illumina
Catalog Number = 16 - 20040536, 96 - 20040537
Website =
Sample Table (Column Headers)
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
LIMS ID (Container)
Built-in
Default = 200
Instruction Notes
Multiline Text
Read Only
Default =
In one well of a new PCR plate, combine the applicable number of 200 ng libraries:
If Enrichment Plexity is 1 then add 7.5 ul of the sample.
If Enrichment Plexity is 3 then add 2.5 ul of each sample to one well with a final volume of 7.5 ul.
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Expand
Well Sort Order = Column
Table Columns - Global Fields
Naming Convention = {InputItemName}
Reagent Kits
Illumina Exome Panel
Supplier = Illumina
Catalog Number = 20020183
Website =
Illumina RNA Prep with Enrichment (L) Tagmentation
Supplier = Illumina
Catalog Number = 16 - 20040536, 96 - 20040537
Sample Table (Column Headers)
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
LIMS ID (Container)
Built-in
Default =
Choose the preheat lid option and set to 100°C
Reaction volume is 25 µl u 95°C for 5 minutes
18 cycles of 1 minute each: u 94°C for the first cycle
Decrease 2°C per subsequent cycle
58°C for 90 minutes
Hold at 58°C for ≤ 24 hours
Step File Placeholders
Log - Manually uploaded
Sample Table
Sample Display Default = Collapse
Well Sort Order = Row
Table Columns - Global Fields
Naming Convention = {InputItemName}
Reagent Kits
Illumina RNA Prep with Enrichment (L) Tagmentation
Supplier = Illumina
Catalog Number = 16 - 20040536, 96 - 20040537
Website =
Sample Table (Column Headers)
Category
Field Name
Field Type
Options
Additional Options and Dropdown Items
Container
Container Name
Built-in
Container
LIMS ID (Container)
Built-in
Decimal Places Displayed = 0
Instruction Notes
Multiine Text
Read Only
Default = Add 23 µl Elution Master Mix to each well.
Thermal Cycler Program Notes
Multiline Text
Read Only
Default =
Choose the preheat lid option and set to 70°C
Reaction volume is 100 µl
Hold at 58°C
Total Samples
Numeric
Default = 0
Decimal Places Displayed = 0
Step File Placeholders
Log - Automatically attached
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Illumina RNA Prep with Enrichment (L) Tagmentation
Default =
Choose the preheat lid option and set to 100°C
Reaction volume is 50 µl
98°C for 30 seconds
14 cycles of:
98°C for 10 seconds
60°C for 30 seconds
72°C for 30 seconds
72°C for 5 minutes
Hold at 10°C
Step File Placeholders
Log - Manually uploaded
Sample Table
Sample Display Default = Collapse
Well Sort Order = Column
Table Columns - Global Fields
Naming Convention = {InputItemName}
Reagent Kits
Agencourt AMPure XP, 5 ml
Supplier = Beckman Coulter
Catalog Number = A63880
Illumina RNA Prep with Enrichment (L) Tagmentation
