Sort By

CloneMiner™ II cDNA Library Construction Kit (Invitrogen™)

The CloneMiner™ II cDNA Library Construction Kit is a second generation CloneMiner™ kit that enables rapid construction of highly representative cDNA libraries without restriction enzyme cloning. This innovative library construction technology combines SuperScript® III Reverse Transcriptase with Gateway® cloning technology, resulting in the discovery of previously unobtainable, full-length clones. The kit also avoids the use of time-consuming, inefficient ligation reactions, making library construction faster and offering better representation.

The CloneMiner™ II cDNA Library Construction Kit ensures:
• High primary titers
• Large average insert sizes
• The highest percentage of full-length genes
• Highly efficient cloning of cDNA to multiple destination vectors without the need of restriction enzyme digestion and ligation

How the CloneMiner™ II Kit Works:

• High yields of full-length cDNA: The CloneMiner™ II kit contains SuperScript® III for generating high cDNA yields. SuperScript® III Reverse Transcriptase (RT), a proprietary mutant of SuperScript® II RT, is active at 50°C and has a half-life of 220 minutes. It also has a point mutation that reduces RNase H activity, thereby decreasing RNA degradation during first-strand synthesis and increasing the percentage of full-length genes.

• Gateway® cloning technology avoids restriction enzyme cloning: Library construction is mediated by Gateway® Technology, a site-specific recombination system that eliminates the use of restriction enzymes and ligase in cloning. Each cDNA insert is flanked by specific att recombination sites (added during the cDNA synthesis steps) that recombine with complementary att sites present in Gateway® donor vectors to create entry clones. Entry clones are subsequently recombined with Gateway® expression vectors to create expression clones, effectively replacing the use of restriction enzymes and ligase. The resulting clones maintain the original orientation and reading frame enabling functional analysis of full-length genes and whole libraries.

How is This Kit Different From the Original CloneMiner™ Kit?
• The new kit incorporates a simplified protocol.
• To ensure high cDNA yields, SuperScript® II Reverse Transcriptase (RT) has been replaced by SuperScript® III Reverse Transcriptase.
• To enable reaction setup with fewer pipetting steps, Gateway® BP Clonase® Enzyme Mix has been replaced by Gateway® BP Clonase® II Enzyme Mix, which contains enzymes and buffer in a single mix.

SuperScript™ Plasmid System for cDNA Synthesis and Plasmid Cloning with Gateway™ Technology (Invitrogen™)

The SuperScript® Plasmid System is designed for synthesis of double-stranded cDNA from a purified mRNA population. The cDNA is suitable for directional cloning into the pSPORT1 vector or the pCMV

• SPORT6 plasmid vector for subsequent generation of cDNA libraries and use of Gateway® Technology. The SuperScript® Plasmid System:
• Results in greater cDNA yields and increased full-length cDNAs with SuperScript® II RT
• Offers a simplified primer-adapter strategy for directional cloning
• Offers one-tube format for first- and second-strand reactions, improving cDNA yields
• Fractionates cDNA using efficient and convenient pre-packed columns
• Ligates inserts into a pre-cut Not I-Sal I vector prepared to minimize background

SuperScript™ Plasmid System with Gateway™Technology and ElectroMAX™ DH10B Competent Cells (Invitrogen™)

The SuperScript Plasmid System is designed for synthesis of double-stranded cDNA from a purified mRNA population. The cDNA is suitable for directional cloning into the pSPORT1 vector or the pCMV

• SPORT6 plasmid vector for subsequent generation of cDNA libraries and use of Gateway Technology. The SuperScript Plasmid System:
• Results in greater cDNA yields and increased full-length cDNAs with SuperScript II RT
• Offers a simplified primer-adapter strategy for directional cloning
• Offers one-tube format for first- and second-strand reactions, improving cDNA yields
• Fractionates cDNA using efficient and convenient pre-packed columns
• Ligates inserts into a pre-cut Not I-Sal I vector prepared to minimize background

Ion AmpliSeq™ Library Kit 2.0 (Ion Torrent™)

The Ion AmpliSeq™ Library Kit 2.0 is designed for rapid preparation of amplicon libraries using Ion AmpliSeq™ panels.

Scalable Multiplex PCR Reactions
The Ion AmpliSeq 2.0 technology enables scalable multiplex PCR reactions from 12- to 24,000-plex in a single well using just 10 ng of starting DNA. Ion AmpliSeq panels and primer pools allow highly multiplexed PCR amplification of thousands of genomic target regions, with superior coverage uniformity and specificity, without the need of a specialized microfluidics PCR platform. In addition, the primers contain proprietary modifications that enable removal of primer sequences during library preparation, for efficient target assessment during sequencing. Multiple primer pools can be used to create overlapping amplicons that enable complete coverage of large targets. Ion AmpliSeq™ Custom Primer Pools are designed via the Ion AmpliSeq™ Designer Tool, available at www.ampliseq.com.

Barcoded Library Preparation
The Ion AmpliSeq Library Kit 2.0 includes reagents for generating amplicons with Ion AmpliSeq primers and preparing libraries from the resulting amplicons. The kit enables the preparation of barcoded libraries using Ion Xpress™ Barcode Adapters 1-96 kits or IonCode™ Barcode Adapters 1-384 kits. Barcoded libraries can be combined and loaded onto a single Ion chip to minimize the sequencing run time and cost and allow for accurate sample-to-sample comparisons.

Easy and Flexible Protocol
The Ion AmpliSeq Library Kit 2.0 uses a plate-based format for easier sample handling and tracking, and for compatibility with automation and high-throughput laboratories. The library preparation uses very small quantities of starting material per PCR. The resulting DNA libraries are ready for downstream template preparation using the automated Ion Chef™ System (which also provides the option of automated library preparation using the Ion AmpliSeq™ Kit for Chef DL8) or the Ion OneTouch™ 2 System followed by sequencing. The intuitive Torrent Suite™ Software and optional Ion Reporter™ Software enables automation of data analysis.

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.

Library Builder™ Whole Transcriptome Core Kit for 5500 Genetic Analysis Systems (Applied Biosystems™)

Library Builder™ Whole Transcriptome Core Kit for 5500 Genetic Analysis Systems is a reagent component of the AB Library Builder™ System. This kit, along with separately purchased Agencourt® AMPure® Beads, allows for the creation of up to 13 RNA whole transcriptome libraries on the Library Builder™.
• Automation – reduce hands on time by 50% or more depending on the number of libraries
• Scalable – prepare up to 13 libraries per run, up to 26 libraries per day
• Integrated – compatible with the SOLiD® 4 System and 5500 Series Genetic Analysis Systems

Labor Saving Automation
The Library Builder™ Whole Transcriptome Core Kit for 5500 Genetic Analysis Systems when used with the AB Library Builder™ System automates the most labor-intensive parts of RNA whole transcriptome library preparation, helping to reduce hands-on time by at least 50%. The Library Builder™ Whole Transcriptome Core Kit for 5500 Genetic Analysis Systems contains enough reagents and materials to process up to 13 samples at a time.

Integrated RNA Library Preparation
The Library Builder™ Whole Transcriptome Core Kit for 5500 Genetic Analysis Systems, along with the AB Library Builder™ System, is an integrated solution comprising a predefined software protocol, plug-and-play cartridge format reagents, and instrument. These kits are all optimized and validated for RNA purification and RNA whole transcriptome library preparation with the 5500 and SOLiD® 4.

To further maximize automation consider using RNA purification protocols with the iPrep™ PureLink® Total RNA Kit, compatible with the Library Builder™ System.

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

Precision ID Library Kit (Ion Torrent™)

The Precision ID Library Kit is designed for rapid preparation of amplicon libraries using HID-Ion AmpliSeq ready-to-use panels, for sequencing on Ion Torrent sequencing system. This kit uses only certain components found in the Ion AmpliSeq™ Library Kit 2.0, that are needed for Precision ID applications, making the Precision ID Library Kit more customized and specific for forensic research use only.

Key features:

Simple protocol
• 96 well plate-based workflow allows easy sample management
• No library amplification with qPCR quantification

Ion AmpliSeq™ HiFi Master Mix
• Enables cleaner amplification for increased coverage uniformity
• Up to 3,072-plex PCR allows larger panel designs

Digestion reagent
• Cleaner digest allows for more efficient downstream sequencing

Benefits for HID applications:

• 1 ng of DNA input is recommended for the Precision ID panels
• Less components to help save time and money by preventing unnecessary waste

Scalable multiplex PCR reactions
The Precision ID Library Kit is built on Ion AmpliSeq chemistry, which enables scalable multiplex PCR reactions from 12- to 3,072-plex in a single well using just 1 ng of starting DNA. Ion AmpliSeq panels and primer pools allow highly multiplexed PCR amplification of thousands of genomic target regions, with superior coverage uniformity and specificity, without the need of a specialized microfluidics PCR platform. In addition, the primers contain proprietary modifications that enable removal of primer sequences during library preparation, for efficient target assessment during sequencing. Multiple primer pools can be used to create overlapping amplicons that enable complete coverage of large targets.

Barcoded library preparation
The Precision ID Library Kit includes reagents for generating amplicons with Ion AmpliSeq primers and preparing libraries from the resulting amplicons. The kit enables the preparation of barcoded libraries using Ion Xpress™ Barcode Adapters 1–96 kits and the IonCode™ Barcode Adapters 1-384 Kit. Barcoded libraries can be combined and loaded onto a single Ion chip to minimize the sequencing run time and cost and allow for accurate sample-to-sample comparisons.

Easy and flexible protocol
The Precision ID Library Kit uses a plate-based format for easier sample handling and tracking, and for compatibility with automation and high-throughput laboratories.

The resulting DNA libraries are ready for downstream template preparation for clonal amplification on Ion Sphere™ particles using the automated Ion OneTouch™ 2 System. The intuitive Torrent Suite Software and HID SNP or STR Genotyper plug-ins enable you to go from extracted DNA to ancestry-informative genotypes in less than two days*.

*Using the Applied Biosystems™ Precision ID NGS System for Human Identification

SOLiD™ Fragment Library Construction Reagents (Applied Biosystems™)

The SOLiD™ Fragment Library Construction Reagents provide a fully optimized set of reagents to generate 10 fragment libraries using the SOLiD™ version 3 standard or express fragment library protocols. This kit replaces S3100105. Additional columns and gels recommended in the SOLiD™ version 3 and 3 plus protocol are available in the SOLiD™ Fragment Library Construction Kits. The SOLiD™ Fragment Library Oligo Kit or an equivalent alternative is also required to generate fragment libraries.

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.

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.

Ion Xpress™ Plus Fragment Library Kit for AB Library Builder™ System (Ion Torrent™)

The Ion Xpress™ Plus Fragment Library Kit for AB Library Builder™ System provides a rapid, flexible, and high-throughput solution for library construction upstream of the semiconductor sequencing workflow. This ready-to-use reagent cartridge simplifies next-generation sequencing by enabling automated DNA fragment library preparation for the Personal Genome Machine® and soon the Proton™ sequencing systems.

This kit contains sample preparation reagents for enzymatic shearing and library construction of 13 DNA libraries for semiconductor sequencing. Use of this reagent cartridge requires the addition of library adapters (sold separately).

This library kit offers:

Simplicity and convenience of automated library preparation upstream of semiconductor sequencing
Superior coverage uniformity for diverse sample types using our proprietary Ion Shear™ Plus chemistry
Lower input requirement providing the ability to make amplification-free libraries from as little as 50 ng of starting DNA
Rapid and flexible workflow producing Ion fragment libraries containing user-selectable and automatically size-selected insert sizes of 100–400 bp in as little as 2 hours
Significant cost savings by removing the need for ancillary shearing devices and size-selection gels
Scalable methodology to meet high-throughput sequencing requirements

With its plug-n-play modular reagent design, this revolutionary kit automates all steps of library creation, including both DNA fragmentation and library size selection, in as little as 2 hours with limited set-up time and minimal subsequent user-intervention. DNA fragmentation is automated with enhanced, proprietary Ion Shear™ DNA fragmentation chemistry to achieve user-selectable inserts of 100–400 bp in length. Size selection of the libraries is performed automatically without the expense or effort of gel-based methods. Alternatively, libraries can be manually size-selected using conventional methods after the automated run. Starting with 50–1000 ng of input DNA, the kit yields significantly higher amounts of library than current manual methods, eliminating the need for PCR amplification (i.e., "amplification-free" libraries) for most sequencing needs.

A Single-Day Workflow: The Next Stage in a Sequencing Revolution
With the ability to prepare up to 13 libraries in a little over 2 hours, the Ion Xpress™ Plus Fragment Library Kit for AB Library Builder™ System is a key step towards automation of the Ion sequencing workflow, making comprehensive analyses of your sample(s) of interest more convenient than ever before. Furthermore, Ion Xpress™ Plus fragment libraries generated using the AB Library Builder™ system exhibit the superb coverage uniformity typically seen with proprietary Ion Shear™ DNA fragmentation chemistry and higher recovery rates than other library construction methodologies. So you can be confident of generating the best data in the shortest time, without the hassle or expense of manual shearing and size selection.

Ion Torrent Sequencing Made Simpler, Faster, and More Affordable
Ion Shear™ Plus DNA fragmentation technology eliminates the need for costly physical shearing devices for most applications and requires as little as 50 ng of sample, accelerating your path to semiconductor sequencing. This flexible technology also provides users the ability to adjust fragmentation and size-selection sizes to accommodate a range of sequencing read lengths depending on their project requirements. This automated library preparation solution provides fast and scalable workflows for sequencing projects requiring low- to high-throughput processing capabilities.

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

Ion Plus Fragment Library Kit for AB Library Builder™ System (Ion Torrent™)

The Ion Plus Fragment Library Kit for AB Library Builder™ System provides a rapid, flexible, and high-throughput solution for library construction upstream of semiconductor sequencing when starting with fragmented DNA. This ready-to-use reagent cartridge simplifies next-generation sequencing by enabling automated DNA fragment library preparation for the Personal Genome Machine® and soon the Proton™ sequencing systems.

This kit contains sample preparation reagents for library construction of 13 DNA libraries for semiconductor sequencing. Use of this reagent cartridge requires additional standard or barcoded library adapters (sold separately).

This library kit offers:

Simplicity and convenience of automated library preparation upstream of semiconductor sequencing
Rapid and flexible workflow automating library creation and size selection in as little as 2 hours starting with pre-fragmented DNA
Lower input requirement providing the ability to make amplification-free libraries from as little as 50 ng starting material
Scalable methodology applicable to automated library preparation systems including the AB Library Builder™ System

With greatly reduced hands-on time, the Ion Plus Fragment Library Kit for AB Library Builder™ System automates preparation of up to 13 standard, paired-end, or barcode fragment libraries in about 2 hours per run, starting with small amplicons or pre-sheared gDNA samples. Size selection of the libraries is performed automatically without the expense or effort of gel-based methods. Alternatively, libraries can be manually size-selected by conventional methods after the automated run.

Starting with 50–1000 ng of input DNA, the kit yields significantly higher amounts of library than current manual methods, eliminating the need for PCR amplification (i.e. "amplification-free" libraries) for most sequencing needs.

A Single-Day Workflow: The Next Stage in a Sequencing Revolution
With the ability to prepare up to 13 libraries in a little over 2 hours, the Ion Plus Fragment Library Kit for AB Library Builder™ System is a key step towards automation of the Ion sequencing workflow, making comprehensive analyses of your sample(s) of interest more convenient than ever before. This modular reagent system allows for the construction of a variety of fragment libraries (i.e., standard and barcoded) depending on the library adapters used (library adapters sold separately).

Ion Torrent Sequencing Made Simpler, Faster, and More Affordable
The Ion Plus Fragment Library Kit for AB Library Builder™ System automates library creation and size selection, eliminating the need for costly time-consuming gel-based size selection, and requires as little as 50 ng of sample, accelerating your path to semiconductor sequencing. This automated library preparation solution provides fast and scalable workflows for sequencing projects requiring low- to high-throughput processing capabilities.

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

SOLiD™ RNA Barcoding Kit, Module 33-48 (Applied Biosystems™)

The SOLiD™ RNA Barcoding Kit, Module 33-48 enables you to increase the power of your studies by cost effectively increasing the numbers of samples analyzed in one SOLiD™ Sequencing run. Compatible with the SOLiD™ Total RNA-Seq Kit and SOLiD™ SAGE™ Kit, the SOLiD™ System barcodes contain unique sequences designed for optimal multiplexing. Module 33-48 contains sixteen different barcodes, selected for uniform melting temperature, low error rate, and orthogonal sequences that are unique in color space. The barcodes are added to the 3' end of the target sequence using a modified P2 adapter. SOLiD™ System barcoding enables the assignment of a unique identifier to templated beads made from a single library. Once the identifiers are assigned, multiple batches of templated beads may be pooled together for emulsion PCR and then sequenced.

Following sequencing, additional rounds of ligation-based sequencing are performed using primer sets complimentary to the barcode. The resulting reads can then be sorted by the barcode and aligned in groups to the reference sequence, enabling the sequencing of multiple libraries on the same slide—maximizing throughput while dramatically reducing costs.
SOLiD™ RNA Barcoding Kits contain PCR primer sets that are designed for use with the SOLiD™ Total RNA-Seq Kit and SOLiD™ SAGE™ Kit. Together, these kits enable multiplex SOLiD™ System sequencing of RNA samples. The SOLiD™ Total RNA-Seq Kit contains two primers:
• The SOLiD™ 5' PCR Primer is identical to the 5’ PCR primer provided in the SOLiD™ Total RNA-Seq Kit, and it contains the SOLiD™ emulsion PCR primer P1 sequence.
• The SOLiD™ 3' (reverse) PCR primers contain the P2 sequence required for SOLiD™ emulsion PCR, along with a unique barcode sequence, and an internal adaptor (IA) sequence necessary for sequencing the barcode (Figure 1). Using these primers in the cDNA amplification step of the SOLiD™ Total RNA-Seq Kit procedure generates barcoded cDNA libraries that can be mixed for multiplex SOLiD™ System sequencing.

SOLiD™ RNA Barcoding Kit, Module 1-16 (Applied Biosystems™)

The SOLiD™ RNA Barcoding Kit, Module 1-16 enables you to increase the power of your studies by cost effectively increasing the numbers of samples analyzed in one SOLiD™ Sequencing run. Compatible with the SOLiD™ Total RNA-Seq Kit and SOLiD™ SAGE™ Kit, the SOLiD™ System barcodes contain unique sequences designed for optimal multiplexing. Module 1-16 contains sixteen different barcodes, selected for uniform melting temperature, low error rate, and orthogonal sequences that are unique in color space. The barcodes are added to the 3' end of the target sequence using a modified P2 adapter. SOLiD™ System barcoding enables the assignment of a unique identifier to templated beads made from a single library. Once the identifiers are assigned, multiple batches of templated beads may be pooled together for emulsion PCR and then sequenced.

Following sequencing, additional rounds of ligation-based sequencing are performed using primer sets complimentary to the barcode. The resulting reads can then be sorted by the barcode and aligned in groups to the reference sequence, enabling the sequencing of multiple libraries on the same slide—maximizing throughput while dramatically reducing costs.
SOLiD™ RNA Barcoding Kits contain PCR primer sets that are designed for use with the SOLiD™ Total RNA-Seq Kit and SOLiD™ SAGE™ Kit. Together, these kits enable multiplex SOLiD™ System sequencing of RNA samples. The SOLiD™ Total RNA-Seq Kit contains two primers:
• The SOLiD™ 5' PCR Primer is identical to the 5’ PCR primer provided in the SOLiD™ Total RNA-Seq Kit, and it contains the SOLiD™ emulsion PCR primer P1 sequence.
• The SOLiD™ 3' (reverse) PCR primers contain the P2 sequence required for SOLiD™ emulsion PCR, along with a unique barcode sequence, and an internal adaptor (IA) sequence necessary for sequencing the barcode. Using these primers in the cDNA amplification step of the SOLiD™ Total RNA-Seq Kit procedure generates barcoded cDNA libraries that can be mixed for multiplex SOLiD™ System sequencing.

GeneRacer™ Kit with SuperScript™ III RT and Zero Blunt™ TOPO™ PCR Cloning Kit for Sequencing (Invitrogen™)

GeneRacer® is an advanced RACE (rapid amplification of cDNA ends) technique that improves the efficiency of amplifying full-length 5´ and 3´ cDNA ends. With the GeneRacer® Kit you can:

• Generate cDNA from transcripts at least 10 kb in length
• Obtain the full-length 5´ end of rare transcripts at fewer than 30 copies per cell
• Clone the full-length 5´ and 3´ ends to construct complete cDNA sequence

The GeneRacer® Kit is available with SuperScript® III Reverse Transcriptase (RT) for improved amplification of the full-length 5´ end from long and complex mRNA. The RNase H portion of SuperScript® III RT has been mutated to avoid cleaving mRNA during cDNA synthesis. This increases the size and yield of cDNA. SuperScript® III RT is more thermostable than wild-type RTs. This enables reverse transcription at higher temperatures, relaxing secondary structure of complex templates, and allowing cDNA synthesis to go to completion.

How GeneRacer® Works

The GeneRacer® Kit ensures that only transcripts containing full-length cDNA ends are amplified (1,2). Figure 1 outlines how the GeneRacer® Kit works. The advanced protocol starts at the RNA level by specifically targeting only 5´ capped mRNA. In subsequent steps the cap is removed and replaced with the GeneRacer® RNA Oligo. During reverse transcription, the GeneRacer® RNA Oligo sequence is incorporated into the cDNA. Only cDNA that is completely reverse transcribed will contain this known sequence. 5´ RACE PCR is then performed using the GeneRacer® 5´ Primer specific to the GeneRacer® RNA Oligo sequence and a gene-specific primer. The result is amplified DNA that contains the full-length 5´ cDNA sequence.

Sensitivity and Length

To demonstrate the ability of the GeneRacer® Kit to capture the full-length 5fi cDNA end, the 5fi ends of genes with known transcriptional start sites were amplified. Starting with total RNA and following the GeneRacer® protocol, both long transcripts (10 kb) and rare messages present at 0.01%, or 30 copies per cell were amplified (Figure 2).