HiSeq® Rapid SBS Kit v2
Reagent Prep Guide
FOR RESEARCH USE ONLY
Reagent Kit Overview
Kit Contents and Storage Requirements
Number of Cycles
Prepare SBS Reagents
Next Steps
Reagent Positions
Technical Assistance
ILLUMINA PROPRIETARY
Part # 15058772 Rev. A
Catalog # FC-402-9003DOC
November 2014
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To perform a rapid run on the HiSeq 2500 or HiSeq 1500, you need at least 1 HiSeq
Rapid SBS Kit v2.
Kit Name
Catalog #
HiSeq Rapid SBS Kit v2 (500 cycles)
FC-402-4023
HiSeq Rapid SBS Kit v2 (200 cycles)
FC-402-4021
HiSeq Rapid SBS Kit v2 (50 cycles)
FC-402-4022
Each kit includes sequencing reagents used on the HiSeq and a set of funnel caps. The
reagents are sufficient for sequencing 1 flow cell.
Kit Component
Quantity
Rapid SBS Kit v2 (Box 1 of 2)
1
Rapid SBS Kit v2 (Box 2 of 2)
1
Set of 8 funnel caps
1
SBS Kit Features
Reagent kits include the following features:
} Reagents are provided in 250 ml bottles that load directly onto reagent racks.
} Reagent labels are color-coded to reduce errors while loading reagents.
} Each reagent contains a barcode for tracking and a human-readable label.
NOTE
Scan the reagent kit ID from box 1 during the run setup steps before loading reagents.
HiSeq Rapid SBS Kit v2 Reagent Prep Guide
3
Reagent Kit Overview
Reagent Kit Overview
Kit Contents and Storage Requirements
HiSeq SBS reagents are shipped in 2 boxes. One box is shipped at room temperature and
the other box is shipped on dry ice. When you receive your kit, promptly store the kit
components at the indicated temperature to ensure proper performance.
NOTE
Reagents are sensitive to light. Store reagents in a dark place.
Reagents are labeled with a 3-letter acronym plus a numeric designator that indicates
the kit size. For example, if you are using CRM-50, the CRM was provided in a 50-cycle
kit.
SBS Kit Box 1
Kit Component
Quantity
Storage
Description
CWM
1
15°C to 30°C
Cleavage Wash Mix
PW1
1
15°C to 30°C
Wash Buffer
USB
2
15°C to 30°C
Universal Sequencing
Buffer
Storage
Description
SBS Kit Box 2
Kit Component
Quantity
IMT
1
-25°C to -15°C
Incorporation Master
Mix
CRM
1
-25°C to -15°C
Cleavage Reagent Mix
USM
1
-25°C to -15°C
Universal Scan Mix
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Part # 15058772 Rev. A
Rapid SBS kits contain a sufficient volume of reagents for performing dual-indexed runs:
} One 500-cycle SBS kit provides reagents for up to 525 cycles of sequencing.
} One 200-cycle SBS kit provides reagents for up to 225 cycles of sequencing.
} One 50-cycle SBS kit provides reagents for up to 74 cycles of sequencing.
} For runs greater than the maximum number of cycles in a kit and to maximize
reagent use for other read lengths, use multiple 50-cycle kits.
The following table lists the number of cycles performed for dual-indexed runs of
various lengths. For dual-indexed paired-end runs, 7 chemistry-only cycles are
calculated in the total number of cycles.
Run Type
Dual-Indexed Paired-End
Dual-Indexed Single-Read
HiSeq Rapid SBS Kit v2 Reagent Prep Guide
Run Length
Run Cycles
Total Cycles
2 x 250
2 x 100
2 x 92
2 x 75
2 x 50
2 x 35
1 x 250
1 x 100
1 x 92
1 x 75
1 x 50
1 x 35
251+8+(7)+8+251
101+8+(7)+8+101
93+8+(7)+8+93
76+8+(7)+8+76
51+8+(7)+8+51
36+8+(7)+8+36
251+8+8
101+8+8
93+8+8
76+8+8
51+8+8
36+8+8
525
225
209
175
125
95
267
117
109
92
67
52
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Number of Cycles
Number of Cycles
Prepare SBS Reagents
SBS reagents are loaded onto the instrument at the beginning of the run. Before setting
up the run, use the following instructions to thaw and inspect reagents.
Thaw Reagents
1
Remove CRM, IMT, and USM from -25°C to -15°C storage.
2
Thaw reagents in a room temperature deionized water bath for about 90 minutes.
Alternatively, thaw reagents at 2°C to 8°C for up to 16 hours.
NOTE
Always replace your gloves after handling CRM.
3
Invert each bottle several times to mix.
4
Inspect the reagents for ice crystals and precipitate. Make sure that the reagents have
thawed completely and precipitate in the USM bottle is cleared.
5
Set CRM, IMT, and USM aside at 2°C to 8°C until you are ready to load them onto
the instrument.
6
Use CWM, PW1, and USB directly from storage.
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Part # 15058772 Rev. A
After you have prepared SBS reagents, you are ready to load them onto the instrument
when prompted by the control software. For reagent loading and sequencing workflow
instructions, see the user guide for your sequencing instrument:
} HiSeq 2500 System User Guide, part # 15035786
} HiSeq 1500 System User Guide, part # 15035788
Make sure that you use the compatible version of cluster kit and control software.
SBS Kit Version
Cluster Kit Version
Control Software
HiSeq Rapid SBS Kit v2
HiSeq Rapid PE Cluster Kit v2
HiSeq Rapid SR Cluster Kit v2
HiSeq Rapid Duo cBot Sample
Loading Kit
HCS v2.2 / RTA v1.18, or later
HiSeq Rapid SBS Kit v2 Reagent Prep Guide
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Next Steps
Next Steps
Reagent Positions
Reagent bottles have color-coded labels that match their positions in the reagent racks.
Load reagents as described in the user documentation for your sequencing instrument.
To prepare for the post-run rinse at the end of a sequencing run, load 25 ml of PW1 or
laboratory-grade water in positions 2 and 4.
Figure 1 SBS Reagent Rack
Table 1 SBS Reagent Positions
Position Reagent
1
2
3
4
5
6
7
8
8
IMT
PW1
USM
PW1
USB
USB
CRM
CWM
Description
Incorporation Mastermix
25 ml of PW1 or laboratory-grade water
Universal Scan Mix
25 ml of PW1 or laboratory-grade water
Universal Sequencing Buffer
Universal Sequencing Buffer
Cleavage Reagent Mastermix
Cleavage Wash Mix
Part # 15058772 Rev. A
For technical assistance, contact Illumina Technical Support.
Table 2 Illumina General Contact Information
Address
5200 Illumina Way
San Diego, CA 92122 USA
Website
www.illumina.com
Email
[email protected]
Table 3 Illumina Customer Support Telephone Numbers
Region
Contact Number
Region
North America
1.800.809.4566
Italy
Austria
0800.296575
Netherlands
Belgium
0800.81102
Norway
Denmark
80882346
Spain
Finland
0800.918363
Sweden
France
0800.911850
Switzerland
Germany
0800.180.8994
United Kingdom
Ireland
1.800.812949
Other countries
Contact Number
800.874909
0800.0223859
800.16836
900.812168
020790181
0800.563118
0800.917.0041
+44.1799.534000
Safety Data Sheets
Safety data sheets (SDSs) are available on the Illumina website at
support.illumina.com/sds.html.
Product Documentation
Product documentation in PDF is available for download from the Illumina website. Go
to support.illumina.com, select a product, then click Documentation & Literature.
HiSeq Rapid SBS Kit v2 Reagent Prep Guide
Technical Assistance
Technical Assistance
*15058772*
Part # 15058772 Rev. A
Illumina
San Diego, California 92122 U.S.A.
+1.800.809.ILMN (4566)
+1.858.202.4566 (outside North America)
[email protected]
www.illumina.com