TruSeq Synthetic Long-Read DNA Library Prep Sequencing Product Insert (15047266 A)

TruSeq Synthetic Long-Read DNA Library Prep Sequencing Product Insert (15047266 A)
Sequencing and Analysis with TruSeq Synthetic Long-Read
DNA Library Prep
FOR RESEARCH USE ONLY
®
®
The Illumina TruSeq Synthetic Long-Read DNA Library
Prep Kit provides a library prep workflow for Human
Genome Phasing to identifying co-inherited alleles and
phasing of de novo mutations, as well as synthetic long-reads
for genome finishing, metagenomics, and de novo sequencing.
The informatics for these two applications is exclusively
enabled by two BaseSpace® apps: TruSeq Long-Read Assembly
and TruSeq Phasing Analysis. Illumina supports sequencing
of TruSeq Synthetic Long-Read DNA libraries on HiSeq® 1000,
2000, 1500, and 2500. The sequencing run must be streamed
to BaseSpace to utilize the corresponding app for data
analysis.
Sample Sheet Preparation
A custom sample sheet is required before starting a
sequencing run with libraries prepared using the TruSeq
Synthetic Long-Read DNA Library Prep Kit. This custom
sample sheet enables correct identification of the sequenced
library using the TruSeq Synthetic Long-Read apps in
BaseSpace. This custom sample sheet can be created using
Illumina Experiment Manager (IEM) v1.8 or later by selecting
the HiSeq FASTQ Only and TruSeq Synthetic Long-Read
DNA sample prep kit options.
cBot™ Workflow
The TruSeq Synthetic Long-Read DNA workflow on the HiSeq
is single indexing. However, TruSeq Synthetic Long-Read
DNA Library Prep is Nextera® based, therefore use the TruSeq
Dual Index Sequencing Primer Box, Paired-End (Catalog # PE121-1003) when performing cluster generation with the TruSeq
PE Cluster Kit v3. Use the TruSeq Dual Index Sequencing
Primer (HP10) in place of the existing Read 1 sequencing
primer (HP6). If you are using TruSeq PE Rapid Cluster kit or
HiSeq PE Cluster Kit v4, no additional primers are required.
For required software version and recipes for each cluster kit,
see the cBot User Guide (part # 15006165).
kit or TruSeq PE Cluster Kit v4, no additional primers are
required.
} HiSeq Software: Sequencing TruSeq Synthetic Long-Read
DNA libraries on HiSeq requires HCS 2.0 or later and a
connection to BaseSpace. Software is available for
download from MyIllumina.
} Workflow: Sequencing TruSeq Synthetic Long-Read DNA
libraries requires paired-end reads of 100–126 (depending
on the SBS kit) and a single eight-cycle Index 1 (i7) Read.
A different sequencing workflow is recommended for each
SBS kit to accommodate reagent usage during run.
Kit
Catalog #
Recipe
TruSeq SBS Kit v3
– 200 cycles
FC-401-3001
100 + 8 (Index 1)
+ 100
TruSeq Rapid SBS Kit
– 200 cycles
FC-401-4001
101 + 8 (Index 1)
+ 101
HiSeq SBS Kit v4
– 250 cycles
FC-401-4001
101 + 8 (Index 1)
+ 101
or
126 + 8 (Index 1)
+ 126
} HiSeq Run Parameters: Select Use BaseSpace for storage
and analysis of TruSeq Synthetic Long-Read DNA
libraries. On the recipe screen, select Custom, then select
Flow Cell Format: Paired End, and then select the number
of cycles for each read.
Sequencing Workflow
} Primers: If sequencing TruSeq Synthetic Long-Read DNA
libraries with the TruSeq PE Cluster Kit v3, use the
sequencing primers HP11 and HP12 within the
TruSeq Dual Index Sequencing Primer Box, Paired-End
(catalog # PE-121-1003). If using TruSeq PE Rapid Cluster
Part # 15047266 Rev. A
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Sequencing and Analysis with TruSeq Synthetic Long-Read DNA Library Prep
View TruSeq Synthetic Long-Read DNA Library Prep online
training courses on the Illumina website:
Analysis Workflow
TruSeq Synthetic Long-Read apps on BaseSpace are required
to analyze TruSeq Synthetic Long-Read DNA library data
generated on the HiSeq system. A sample sheet generated by
IEM v1.8 or higher is required for analysis in BaseSpace. For
more information, see the TruSeq Long-Read Assembly User
Guide (part # 15055850) or TruSeq Phasing Analysis User Guide
(part # 15055852).
1
Go to support.illumina.com.
2
Select Sequencing.
3
Select Kits & Reagents.
4
Select TruSeq Synthetic Long-Read DNA Library Prep
Kit to open the support page.
5
Select Training.
Software Requirements
Technical Assistance
Software upgrades might be required to run TruSeq Synthetic
Long-Read DNA libraries. To download the software version
required for your Illumina instrument, go to
support.illumina.com/sequencing/downloads.ilmn:
} IEM: IEM 1.8 or later
} cBot software (if needed): cBot 1.4 software or later (v3) or
cBot 2.0.1 or later (v4); the recipe installer can also be
found under this listing.
} HCS software for HiSeq: HCS 2.0/RTA 1.17 or later
For questions, go to the Support tab on www.illumina.com. If
you do not find the information you need there, contact
Illumina Technical Support by email or phone.
Supporting Documentation and Training
To download the latest version of the documentation, go to
www.illumina.com/support.
Document Title
Part #
BaseSpace User Guide
15044182
cBot User Guide
15006165
HiSeq 1000 User Guide
15023355
HiSeq 1500 User Guide
15035788
HiSeq 2000 User Guide
15011190
HiSeq 2500 User Guide
15035786
IEM TruSeq Synthetic Long-Read DNA Quick
Reference Card
15056316
Illumina Experiment Manager User Guide
15031335
TruSeq Long-Read Assembly User Guide
15055850
TruSeq Phasing Analysis User Guide
15055852
TruSeq Synthetic Long-Read DNA Library Prep
Guide
15047264
TruSeq Synthetic Long-Read DNA Library Prep
Experienced User Card and Lab Tracking Form
15047265
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Copyright and Trademarks
© 2014 Illumina, Inc. All rights reserved.
Illumina, 24sure, BaseSpace, BeadArray, BlueFish, BlueFuse, BlueGnome,
cBot, CSPro, CytoChip, DesignStudio, Epicentre, GAIIx, Genetic Energy,
Genome Analyzer, GenomeStudio, GoldenGate, HiScan, HiSeq, HiSeq X,
Infinium, iScan, iSelect, ForenSeq, MiSeq, MiSeqDx, MiSeq FGx,
NeoPrep, Nextera, NextBio, NextSeq, Powered by Illumina, SeqMonitor,
SureMDA, TruGenome, TruSeq, TruSight, Understand Your Genome,
UYG, VeraCode, verifi, VeriSeq, the pumpkin orange color, and the
streaming bases design are trademarks of Illumina, Inc. and/or its affiliate(s)
in the U.S. and/or other countries. All other names, logos, and other
trademarks are the property of their respective owners.
Part # 15047266 Rev. A
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