Shop All Library Preparation Kits for Sequencing

Ion 510™ Food Protection Chip Kit (Thermo Scientific™)

Simplify sample tracking and sequencing using this Chip Kit as part of a complete Thermo Scientific™ NGS Workflow for Food Authenticity.

Ion AmpliSeq™ Transcriptome Mouse Gene Expression Kit (Ion Torrent™)

The Ion AmpliSeq Transcriptome Mouse Gene Expression Assay allows simultaneous gene expression measurement of over 20,000 mouse RefSeq genes in a single assay. Providing the ability to work directly with low input levels of total RNA, the Ion AmpliSeq Transcriptome Mouse Gene Expression Assay is ideal for those interested in obtaining comprehensive and highly sensitive gene expression measurements from a wide range of RNA sources. This particular kit enables manual library preparation with throughput flexibility. For automated production, see the Ion AmpliSeq Transcriptome Mouse Gene Expression Panel, Chef-Ready Kit.

The Ion AmpliSeq Transcriptome Mouse Gene Expression Assay offers:
• Flexible, straightforward library preparation—start with as little as 10 ng of total RNA from tissues, cells, or other sources
• Fast gene expression profiling with simple workflow—go from RNA to data in <2 days with ~45 minutes of total hands-on time with the automated workflow of the Ion GeneStudio S5 and Ion Chef systems
• A complementary software plug-in for automated sequencing analysis that provides gene quantitation and reporting as well as quality control metrics
• Compatibility with Ion Xpress Barcode adapters, allowing for multiplexed sequencing

Leveraging the speed and simplicity of Ion AmpliSeq technology, this ultrahigh-multiplex PCR enrichment approach enables rapid gene expression profiling with minimal hands-on time. This research method provides a faster, simpler, and lower-cost sequencing method that is highly correlated to whole-transcriptome RNA sequencing.

Straightforward library preparation and multiplex sequencing with manual kit
Starting with as little as 10 ng of total RNA, libraries can be prepared in approximately seven hours with only one hour of hands-on time. Total RNA is first processed with the SuperScript VILO cDNA Synthesis Kit (not included), and the resulting cDNA is amplified using Ion AmpliSeq technology to accurately maintain expression levels of all genes.

The resulting libraries are ready for downstream template preparation using the Ion OneTouch 2 or Ion Chef System and sequencing on the Ion GeneStudio S5 or Ion Proton System. Libraries can be barcoded for multiplexing, providing flexibility, reduced cost, and increased throughput, with the recommended 16 libraries per Ion 550 Chip and 8 libraries per Ion 540 or Ion PI chip. Other multiplexing options are also possible, depending on the experimental design and desired level of sensitivity.

Ion Library Equalizer™ Kit (Ion Torrent™)

The Ion Library Equalizer™ Kit is a simple and seamless bead-based solution replacing the need for library quantification and library dilutions for library normalization as required for any next generation sequencing workflow. The Ion Library Equalizer™ Kit helps reduce labor, reagent costs, and further simplifies the Ion semiconductor sequencing workflow for high sample-throughput or barcoded projects, as well as single or low sample number projects.

The kit workflow consists of three simple steps:

1) Library amplification with Equalizer™ Primers
2) Library capture onto Equalizer™ Beads
3) Heat elution of the normalized library using a specially formulated Equalizer™ Elution Buffer

Simple Workflow
• Seamlessly integrates into your existing library preparation workflow
• Reduced pipetting compared to the Agilent 2100 Bioanalyzer®-based workflow
• Faster workflow in high-throughput applications-no need for DNA quantification, calculations, and dilution of DNA libraries
• Minimizes library-to-library variability

Cost Effective
• No additional capital equipment required
• Less expensive than Agilent 2100 Bioanalyzer®-based workflow (up to 10 samples)
• Reduced AMPure® XP reagent usage compared to Agilent 2100 Bioanalyzer®-based workflow
• Long shelf life: up to 18 months
• Convenient 96-reaction kit helps reduce ordering frequency

Flexible
• Compatible with all Ion DNA library kits including Ion AmpliSeq™ DNA panels, Ion fragment library kits, and Ion Xpress™ Barcode adaptors
• Use with a single sample or in a 96-well plate format
• Compatible with 3rd party automated liquid handling systems for integrated library preparation and normalization

Includes:
The Ion Library Equalizer Kit includes the necessary reagents to complete 96 normalization reactions.

Required for Workflow:
Ion AmpliSeq™ Library Kit 2.0
Ion AmpliSeq™ Ready-to-Use Panel or Custom Panel designed via the Ion AmpliSeq™ Designer (www.ampliseq.com)
--OR--
Ion Plus Fragment Library Kit
--OR--
Ion Xpress™ Plus Fragment Library Kit

Optional:
Ion Xpress™ Barcode Adapters Kits

Ion AmpliSeq™ Kit for Chef DL8 (Ion Torrent™)

The Ion AmpliSeq™ Kit for Chef DL8 is designed for use with the Ion Chef™ System for automation of Ion AmpliSeq library construction. The kit offers:

• Simplest workflow for targeted sequencing library preparation using 1 or 2 pool Ion AmpliSeq panel designs*
• Low input
• Easy-to-use cartridge format with reagent barcoding for full reagent tracking
• Proven performance of Ion AmpliSeq technology, compatible with DNA extracted from formalin-fixed, paraffin-embedded tissue and other challenging sources

*See www.AmpliSeq.com for custom 1-2 pool panels. Note that some panels (e.g., Transcriptome, Cancer Hotspot, Oncomine BRCA, Pharmacogenomics) have been prepared in an Ion Chef-ready kit by incorporating the respective ready-made primer pool with the DL8 kit.

Leveraging the automation capabilities of the Ion Chef System, the Ion AmpliSeq Kit for Chef DL8 enables the reproducible preparation of Ion AmpliSeq amplicon libraries, with less than 15 minutes of hands-on time and only 3 liquid transfer steps. Compatible with 1- and 2-pool Ion AmpliSeq panel designs, the kit automates the preparation of libraries from 8 samples in a single run, delivering a single tube of pooled libraries ready for downstream templating and sequencing. The kit provides sufficient material for the preparation of 32 libraries (4 Ion Chef runs).

Utilizing plug and play, pre-packaged, single-use cartridges and PCR plates for a fully automated workflow, users of all experience levels will find library preparation straightforward with minimal effort. Supplied with barcodes conveniently dried-down in 96-well plates, multiplexing samples is easy and requires no additional pipetting steps.

More information on Ion AmpliSeq technology >

Ion Plus Fragment Library Kit (Ion Torrent™)

As an integral component of the Personal Genome Machine™ sequencing platform, the Ion Plus Fragment Library Kit is designed to produce high quality DNA libraries via multiple possible workflows. The economically priced Ion Plus Fragment Library Kit provides low cost sample preparation, bringing next generation sequencing to every lab. This revolutionary kit allows completion of library preparation in as little as 2 hours for gDNA and amplicon libraries when combined with either physical shearing methods or our proprietary Ion Shear™ enzymatic fragmentation technology (Ion Xpress Plus Library Kit, Cat. No. 4471269), thus saving time and cost. With significantly higher yields than other library construction techniques, high quality libraries can be attained with as little as 100 ng input DNA. In addition, this highly efficient library construction process enables the creation of “amplification-free" libraries from <1 ug input material. This kit is also extremely flexible in that it is compatible with multiple physical shearing and library size selection methods.

The Ion Plus Fragment Library Kit contains sample preparation reagents for library construction of up to 20 DNA libraries (depending on input DNA type and amount) for semiconductor sequencing. Store components at -20°C.

The Ion Plus Fragment Library Kit offers you:
• Lower input requirement with as little as 100 ng starting material
• Rapid and flexible workflow with Ion fragment libraries containing variable insert sizes with a choice of mix-and-match workflows
• Scalable methodology applicable to automated library preparation systems including the AB Library Builder™ System

A Single-Day Workflow: The Next Stage in a Sequencing Revolution
With a quick turnaround time the Ion Plus Fragment Library Kit shortens the Ion sequencing workflow to a single workday, making comprehensive analyses of your samples possible in less time than ever before. Furthermore, the Ion Plus Fragment Library Kit is compatible with a variety of DNA shearing and library size-selection methodologies allowing you to tailor your workflow to suit your project needs.

For research use only. Not intended for human or animal therapeutic or diagnostic use.

MuSeek Index Set 1, Illumina compatible (Thermo Scientific™)

Thermo Scientific MuSeek Index Set 1, Illumina™ compatible, is designed for generation of indexed genomic DNA libraries for sequencing on the Illumina MiSeq™ and HiSeq™ instruments. The set provides 4 indexed P5 adapters and 6 MuSeek indexed P7 adapters. This enables pooling of up to 24 dual-indexed libraries and conducting multiplexed sequencing analysis. The MuSeek Index Set 1, in combination with MuSeek Library Preparation kit, Illumina™ compatible, allows fast and simple indexed library construction from 50 ng of sample DNA, eliminating the need for physical shearing, end conversion and adapter ligation steps.

Highlights

• Enables sequencing of up to 24 dual-indexed fragment libraries in a single sequencing run.

Applications

• Indexed DNA fragment library preparation for next generation sequencing.

Includes
• MuSeek Indices M501-4
• MuSeek Indices M701-6

IonCode™ Barcode Adapters 1-384 Kit (Ion Torrent™)

The IonCode Barcode Adapters 1-384 Kit provides a set of 384 unique barcode adapters specifically designed for optimal performance with the Ion GeneStudio S5, Ion PGM, and Ion Proton semiconductor sequencers. When used in combination with Ion Barcode Adapters 385-768 and Ion AmpliSeq Library Kit PLUS or Ion AmpliSeq Library Kit 2.0, this kit enables users to pool up to 768 amplicon libraries and then conduct multiplexed sequencing analysis, simplifying the Ion semiconductor sequencing workflow for a wide range of applications, including targeted enrichment.

Key product features:

• Enables multiplexing of numerous amplicon library samples on a single sequencing chip with the use of robust molecular barcodes
• Both sequence- and flow-optimized for equal representation of all barcodes in a pool and more economical multiplexed sequencing runs
• Robust error correction built in by design for added confidence in sample identification

Multiplexing with barcode adapters enables higher throughput
Multiplexing with barcoded libraries leads to more cost effective runs by permitting up to 768 samples per run, significantly decreasing the cost and handling requirements of sequencing. Each barcode was individually tested to minimize representation bias.

Sequence- and flow-optimized for increased performance and efficiency
Barcode adapters contain sequences that are optimized to provide equal representation of all barcodes in a pool and require minimal number of flows to interrogate, resulting in more economical multiplexed sequencing runs.

Robust error correction built in by design
Built-in error correction prevents the possibility of incorrect sample assignment.

5500 SOLiD™ Fragment Library Barcode Adaptors 1-16 (Applied Biosystems™)

The 5500 SOLiD™ Fragment Library Barcode Adaptor Kit provides a complete set of 96 unique barcode adaptors and primers for cost-effective next generation sequencing of fragment libraries on the 5500 Series Genetic Analysis Systems. When used in combination with the 5500 SOLiD™ Fragment Library Core Kit, the 5500 SOLiD™ Fragment Library Barcoding Kit enables users to pool up to 96 fragment libraries prior to emulsion PCR and then conduct multiplexed sequencing analysis, simplifying the next generation sequencing workflow for a wide range of applications, including targeted enrichment

Key Product Features:
• Efficient multiplexing—enables multiplexing of up to 96 fragment or paired-end library samples on a single flow chip with the use of robust molecular barcodes.
• Cost effective runs—allows more samples per run, significantly decreasing the cost and handling requirements of ePCR, enrichment, and deposition.
• Consolidated 5500 SOLiD sampling—enables pooling of samples prior to emulsion PCR (ePCR) and sequencing on the 5500 Series Genetic Analysis Systems.
• Versatile sequencing—substantially benefits applications such as whole genome sequencing of small genomes or targeted resequencing of enriched samples.

For Research Use Only. Not for use in diagnostics procedures.

Ion Ampliseq™ Cancer Hotspot Panel v2 Chef-Ready Kit (Ion Torrent™)

The Ion Ampliseq™ Cancer Hotspot Panel v2 Chef-Ready Kit combines a single pool of primers with the reagents necessary to prepare libraries of frequently mutated regions in human cancer genes on the Ion Chef™ System. This automated workflow utilizes the same chemistry as the manual library construction workflow. It provides equivalent or better performance with exceptional reproducibility, on-target reads, and sample balance. Automated production of libraries using the Ion Chef System reduces hands-on time to less than 15 minutes.

The Ion Ampliseq Cancer Hostpot Panel v2 Chef Ready Kit consists of the following components:
• Ion Ampliseq™ Cancer Hotspot Research Panel Primers
• Ion Ampliseq™ Kit for Chef DL8

The simple sample-to-data workflow can be executed in less than two days and offers the following features and benefits:
• Production of high-quality DNA libraries using the Ion Chef System
• Proven performance of Ion Ampliseq™ technology, FFPE compatibility, uniform coverage for efficient sequencing, accurate variant calling, and improved variant detection via Torrent Suite™ Software
• Ability to scale and multiplex on Ion™ chips based on desired throughput needs, driving cost savings and efficiency
• Simplified workflow and reduced hands-on time when used with the Ion Ampliseq™ Direct FFPE DNA Kit
• Low input amount of only 1 ng per sample

Additional mutational content for advancement of cancer research studies
The Ion AmpliSeq Cancer Hotspot Panel v2 is designed to amplify 207 amplicons covering approximately 2,800 COSMIC mutations from 50 oncogenes and tumor suppressor genes. The panel should be used with the Ion AmpliSeq™ Library Kit 2.0.

Minimal hands-on time
Leveraging the automation capabilities of the Ion Chef System, the included Ion AmpliSeq Kit for Chef Kit DL8 enables the reproducible preparation of Ion AmpliSeq amplicon libraries, with less than 15 minutes of hands-on time and only 3 liquid transfer steps. Compatible with 1- and 2-pool Ion AmpliSeq panel designs, the kit automates the preparation of libraries from 8 samples in a single run, delivering a single tube of pooled libraries ready for downstream templating and sequencing. The kit provides sufficient material for the preparation of 32 libraries.

Simplicity and ease-of-use of Ion solutions
Utilizing plug and play, pre-packaged, single-use cartridges and PCR plates for a fully automated workflow, users of all experience levels will find library preparation straightforward with minimal effort. Supplied with barcodes conveniently dried-down in 96-well plates, multiplexing samples is easy and requires no additional pipetting steps.

Collibri™ 3´ mRNA Library Prep Kit for Illumina™ Systems (Invitrogen™)

The Invitrogen Collibri 3’ mRNA Library Prep Kit is designed for robust construction of cDNA libraries for strand-specific RNA sequencing on Illumina next-generation sequencing (NGS) platforms. It supports library preparation from polyadenylated RNA in ~4.5 hours. The protocol generates one fragment per transcript, resulting in accurate gene expression values. Sequences obtained are close to the 3’ ends of the transcripts, so the kit is suitable for varying-quality RNA samples, including formalin-fixed paraffin-embedded (FFPE) samples. The majority of inserts are greater than 75 bp in size, corresponding to final library fragment sizes ≥200 bp. Multiplexing of libraries can be carried out using up to 96 single-indexed primers.

For convenience, the Collibri 3’ mRNA Library Prep Kit provides color-coded components for visual tracking of library preparation progress. Inert dyes in the reagents provide visual confirmation of additions and mixing, but do not interfere with enzymatic reactions and do not compromise library prep and sequencing results.

Ion Xpress™ Plus Fragment Library Kit (Ion Torrent™)

The Ion Xpress™ Plus Fragment Library Kit provides a rapid and flexible enzymatic-based library construction methodology for the Personal Genome Machine™ sequencing system workflow. Employing enhanced proprietary Ion Shear™ DNA fragmentation chemistry, this revolutionary kit allows completion of library preparation in as little as 2 hours for gDNA and amplicon libraries, removing the need for a physical shearing device to save time and cost. With significantly higher yields and lower bias than other library construction techniques, superior coverage uniformity is attained with as little as 100 ng input DNA. In addition, highly efficient library construction enables the creation of "amplification-free" libraries from 1 µg input material. This kit allows users to modulate an appropriate fragment insert size depending on desired read length and application.

The Ion Xpress™ Plus Fragment Library Kit contains sample preparation reagents for enzymatic shearing and library construction for up to 20 DNA libraries (depending on input DNA type) for semiconductor sequencing. Store components at -20°C.

The Ion Xpress™ Plus Fragment Library Kit offers you:
• Superior coverage uniformity for diverse sample types using our proprietary Ion Shear™ Plus chemistry
• Lower input requirement with as little as 100 ng starting material
• Rapid and flexible workflow with Ion fragment libraries containing variable insert sizes in as fast as 2 hours
• Significant cost savings by removing the need for ancillary shearing devices
• Scalable methodology applicable to automated library preparation systems including the AB Library Builder™ System

A Single-Day Workflow: The Next Stage in a Sequencing Revolution
With a turnaround time of as little as 2 hours, the Ion Xpress™ Fragment Library Kit shortens the Ion sequencing workflow to a single workday, making comprehensive analyses of your sample(s) of interest possible in less time than ever before. Furthermore, Ion Xpress™ Plus fragment libraries exhibit greater coverage uniformity and higher recovery rates across diverse sample types than other library construction methodologies, so you can be confident of generating the best data in the shortest time.

Ion Torrent Sequencing Made Simpler, Faster, and More Affordable
Ion Shear™ Plus DNA fragmentation technology, included in the Ion Xpress™ Plus kit, obviates the need for costly physical shearing devices for most applications and requires as little as 100 ng of sample, accelerating your path to semiconductor sequencing. This flexible technology provides users the ability to adjust the resulting DNA fragment size to accommodate a range of sequencing read lengths depending on their project requirements. This technology is also amenable to automated library preparation systems such as the AB Library Builder™ System to provide quick answers when you need them most. Ion Shear™ Plus DNA fragmentation technology is not included in the Ion Plus kit. Use the Ion Plus kit if physical shearing methods are preferred.

For Research Use Only. Not intended for human or animal therapeutic or diagnostic use.

Ion AmpliSeq™ HD Library Kit (Ion Torrent™)

The Ion AmpliSeq HD Library Kit is designed for generation of amplicon libraries for use as part of the Ion AmpliSeq HD sequencing workflow. The kit is compatible with custom panels created using the Ion AmpliSeq Designer tool (www.ampliseq.com) and sequenced using an Ion GeneStudio S5 Series instrument. Ion AmpliSeq HD technology enables scalable polymerase chain reactions (PCRs) from 12- to 500-plex in a single well (1000-plex in two wells), with sample input amounts as small as 1 ng and a level of detection down to 0.1% depending on the sample source.

Key product features include:
• Custom design creation
• Ultra-high sensitivity detection
• Dual-barcoded sample multiplexing
• Bi-directional sequencing

Custom design creation
The Ion AmpliSeq Designer tool can now be used to create custom designs for use with Ion AmpliSeq HD technology. The designs are made-to-order and allow the user the flexibility of creating region designs, hotspot designs, or fusion designs. These designs are one- or two-pool designs and recommended for use with samples obtained from cfDNA , cfRNA, cfTNA, or FFPE.

Ultra-high sensitivity detection
Ion AmpliSeq HD technology is capable of ultra-high levels of detection down to 0.1%. These detection levels have been observed for cfDNA controls with known variants using an input amount of 20 ng. Levels of detection for other sample types, such as FFPE, will vary.

Dual-barcoded sample multiplexing
The Ion AmpliSeq HD Library Kit includes reagents for generating amplicons using Ion AmpliSeq HD primers, which are used to prepare libraries for sequencing. The libraries are dual-barcoded at the 5’ and 3’ ends using the Ion AmpliSeq HD Dual Barcode Kit 1-24, which enables the multiplexing of up to 24 samples in a single chip.

Bi-directional sequencing
The Ion AmpliSeq HD Library Kit also provides sequencing information from both strands. This enables better error correction and higher confidence in the quality of your sequence data.

Oncomine™ Immune Response Research Assay (Ion Torrent™)

The Oncomine™ Immune Response Research Assay is a targeted gene expression assay designed for the Ion™ next-generation sequencing (NGS) platform. This pan-cancer gene expression assay is designed to interrogate the tumor microenvironment to enable mechanistic studies and identification of predictive biomarkers for immunotherapy in retrospective or prospective clinical trial research cohorts. The assay is optimized to measure the expression of genes involved in tumor-immune interactions, including the low-expressing genes involved in inflammatory signaling.

This assay contains the reagents for manual library construction and a single pool of primers used to perform multiplex PCR for preparation of amplicon libraries from formalin-fixed paraffin-embedded (FFPE) samples. The assay enables quantitative evaluation of the expression of markers associated with different leukocyte subsets, antigen presentation, checkpoint pathways, and tumor progression. This 400-gene assay is supported with a user-friendly informatics workflow and demonstrates high sensitivity for low-expressing transcripts derived from FFPE samples (request a gene list). The Oncomine Immune Response Research Assay leverages Ion AmpliSeq™ technology to deliver results from as little as 10 ng of total RNA, enabling robust performance from challenging FFPE samples.

Assay workflow
The Oncomine Immune Response Research Assay manual workflow features high sample multiplexing on Ion S5™ and Ion PGM™ sequencing systems. The entire workflow, from sample extraction to results, can be completed in less than 48 hours. Note: An alternate assay is available (Cat. No. A32928) for the preparation of Ion AmpliSeq libraries in automated mode through use of an Ion Chef™ System.

Assay performance
• High repeatability and sensitive detection of gene expression across different types of solid tumors
• Low sample input requirement (10 ng of total RNA material)
• High sensitivity detection of low-expressing signaling molecules such as interleukins and interferon gamma
• High sample multiplexing with flexibility to run 8 samples per single run
• Efficient workflow that offers sample-to-results in less than 48 hours
• Automated and seamlessly integrated analysis solution
• Interpretation of immune surveillance and tumor progression mechanism with functional annotation of genes and pathways

Informatics
The Oncomine Immune Response Research Assay comes with optimized informatics and visualization software that minimizes the complexity of data analysis. The analysis workflow consists of a data exploration component automated by the Immune Response Torrent Suite™ Plug-in and a two-group differential analysis provided by the Affymetrix™ Transcriptome Analysis Console (TAC). Together these tools offer a streamlined process for common gene expression analyses and exportable data files that are amenable to further exploration with other third-party software. Specifically, these tools provide:
• QC metrics and expression of housekeeping genes to qualify data
• Hierarchical clustering of samples with user-defined gene sets or by all genes on the panel
• Visualization with PCA, heatmaps, and gene expression distribution plots
• Sample correlation using rich correlation plots
• Fold-change estimates in easy-to-use tabular data format (TAC)
• Volcano plots to visualize differentially genes (TAC)

AgriSeq™ HTS Library Kit (Applied Biosystems™)

The Applied Biosystems AgriSeq HTS Library Kit is specifically designed for high-throughput preparation of amplicon libraries for targeted genotyping-by-sequencing (GBS) applications in agrigenomics.

Key product features include:
• Exceptional uniformity of multiplexed target amplification generates robust, reproducible results
• Simplified workflow enables high-throughput sample processing
• Convenient configuration contains library normalization reagents and volumes sufficient for automated platforms

Excellent data quality
AgriSeq library preparation has been optimized for targeted GBS of panels ranging from 100–5000 amplicons. With a highly efficient and specific multiplex technology, AgriSeq amplification can generate libraries with high uniformity (87.2–99.3%), percent on-target reads (85.7–99.7%), and overall genotype call rates (87.5–99.7%) based on a set of nine recently designed panels ranging in size from 62–3000 amplicons across seven different species. The figure below shows an average of 98.2% uniformity for a 3000-amplicon panel across 192 samples (i.e., 98% of bases are covered by at least 0.2X of the average read depth). Even coverage is significant in that it reduces the amount of sequencing required to achieve consistent genotype calls, thus reducing the sequencing cost per sample. In comparison, weaker multiplexing performance is reported with the competitor technology TruSeq, which provides only 88% average uniformity and a reported 1536 maximum multiplexing capability (see reference).

High throughput sample processing
The AgriSeq Library workflow (see figure below) can be performed in either 96- or 384-well plates for high-throughput sample processing using as little as 10 ng of DNA input per reaction. The workflow has been optimized to minimize sample handling and ancillary reagent usage through the processing of pooled samples. This allows the workflow to be completed with minimal pipetting steps, helping to reduce the potential for error, as well as simplifying automation setup on common third party platforms such as Tecan or Hamilton systems. Standard data analysis processing includes automated quality assessment, alignment, and genotype calling using the included Torrent Suite software and Variant Caller plug-in without the need for a bioinformatician, expensive data storage, or third party programs.

Conveniently packaged containing library normalization reagents & volumes for automation
The AgriSeq HTS Kit is offered in large pack sizes (960 or 9600 reactions) to accommodate high throughput agrigenomics projects while minimizing the cost and waste of unnecessary packaging. The AgriSeq HTS Kit includes not only the reagents for library preparation, but all of the components required for library normalization to help achieve consistent results between samples, and sufficient overage has been provided to accommodate automated workflows. With a low price per sample exclusively available for agricultural customers, AgriSeq targeted GBS is an economical way to generate high-throughput genotyping data to advance your breeding programs.

Ion Total RNA-Seq Kit for AB Library Builder™ System (Ion Torrent™)

The Ion Total RNA-Seq Kit for AB Library Builder™ System combines optimized reagents and protocols for automated preparation of representative cDNA libraries for strand-specific RNA sequencing of all types of RNA species on the Ion Personal Genome Machine (PGM™), Ion Proton™, and Ion GeneStudio™ systems. Either small RNA (such as microRNA) or whole transcriptome RNA samples can be prepared for next-generation sequencing.

The Ion Total RNA-Seq Kit for AB Library Builder™ System contains the same performance advantages as the manual Ion Total RNA-Seq Kit v2 with the convenience of protocol automation. The gel size selection step in the small RNA workflow and the filter cleanup steps have all been replaced with a magnetic bead-based method that reduces the total reaction time to approximately 6 hours. This automated workflow eliminates the hassles associated with manually producing RNA libraries for RNA expression analysis applications. This instrument and reagent combination provides researchers the ability to produce ~36 libraries in a typical work day for medium to high throughput sequencing labs.

The Ion Total RNA-Seq Kit for AB Library Builder™ System is designed to make cDNA library preparation for semiconductor sequencers simple, fast, and flexible. It can be used to generate a representative cDNA library from any type of RNA sample. The Ion Total RNA-Seq Kit for AB Library Builder™ System provides an automated total RNA library solution employing a common workflow for the discovery of small RNAs and isoforms, coding RNA, noncoding RNA, and alternative splice variants.

Additional Features of the New Ion Total RNA-Seq Kit for AB Library Builder™ System:

• Automated workflow—magnetic bead-based purification simplifies automation of library steps post RNA fragmentation
• Greater accuracy—SuperScript® III and Platinum® PCR SuperMix High Fidelity added for highest template fidelity
• Barcode compatible—works with Ion Xpress™ RNA-Seq Barcode 01-16 Kit for multiplexing

As with the manual Ion Total RNA-Seq Kit v2, the Ion Total RNA-Seq for AB Library Builder™ System:

• Preserves strand information—all mapped reads are aligned in the direction of transcription relative to the chromosomal strand
• Allows you to choose your workflow—interrogate either small RNA or the whole transcriptome
• Allows you to analyze any type of RNA—supports small RNA, rRNA depleted total RNA, and poly(A) RNA

Small RNA Analysis
During small RNA library construction, the 3' and 5' adaptors are attached directionally and simultaneously. As a by-product of this step, an adaptor product may be formed without an RNA insert. This byproduct will amplify during first strand synthesis and PCR. If not removed, >50% of the reads will be the adaptor dimer. Historically, the only way to separate the adaptor dimer from the wanted small RNA-containing library fragments has been through gel size selection. The Ion Total RNA-Seq Kit for AB Library Builder™ System uses proprietary technology to inhibit cDNA synthesis of the adaptor byproduct, thus allowing cDNA separation with magnetic bead-based technologies. Total reaction time has been reduced to approximately 3 hours on the instrument. A separate Magnetic Bead Cleanup Module is included with the kit and additional modules may be purchased separately if needed.

Start with total RNA containing as little as 5–100 ng of miRNA, or RNA enriched for small RNA containing 1–100 ng of miRNA. The small RNA protocol provides guidance on whether to start with total RNA or RNA enriched for small RNA, based on the small-RNA content of your sample. Small RNA enrichment protocols are also provided.

Whole Transcriptome Analysis
The whole transcriptome protocol enables construction of strand-specific libraries in approximately 5 hours. Starting with as little as 100 ng of total RNA, construct a library from 1 ng of poly(A) RNA or 25 ng of rRNA-depleted RNA following the RNA enrichment and library generation protocols provided in the manual. Because the libraries are not limited to cDNA derived only from poly(A) RNA, Ion Total RNA-Seq Kit libraries support a more thorough investigation of transcriptome complexity, capable of characterizing known and undocumented transcripts, including alternative splice variants, fusion transcripts, and SNPs.

Preserve Strand Information
Unlike methods that ligate adapters to double-stranded cDNA, the Ion Total RNA-Seq Kit utilizes proprietary Ambion® technology to attach the adapters in a directional manner that preserves strand information in the resulting libraries. In addition, both the 3' and 5' adapters are attached simultaneously, reducing ligation and clean-up steps.Preserving strand orientation during library construction helps enable more accurate determination of the structure and expression level of transcripts, and can aid in the discovery of novel transcription regions from both the positive and negative genomic strands.