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SuperScript™ III First-Strand Synthesis System Invitrogen™

The SuperScript® III First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA. RNA targets from 100 bp to >12 kb can be detected with this system. The amount of starting material can vary from 1 pg–5 µg of total RNA. SuperScript® III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme is used to synthesize cDNA at a temperature range of 42–55°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript® III RT is not significantly inhibited by ribosomal and transfer RNA, it may be used to synthesize first-strand cDNA from a total RNA preparation.

Using the SuperScript® III First-Strand System
cDNA synthesis is performed in the first step using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. In the second step, PCR is performed in a separate tube using primers specific for the gene of interest. For the PCR reaction, we recommend one of the following DNA polymerases: Platinum® Taq DNA Polymerase provides automatic hot-start conditions for increased specificity up to 4 kb, Platinum® Taq DNA Polymerase High Fidelity provides increased yield and high fidelity for targets up to 15 kb, and Platinum® Pfx DNA Polymerase provides maximum fidelity for targets up to 12 kb.

TaqMan™ MicroRNA Reverse Transcription Kit Applied Biosystems™

The Applied Biosystems® TaqMan® MicroRNA Reverse Transcription Kit provides the necessary components for 1000 reactions of optimal performance in TaqMan® MicroRNA Assays. Components of this kit are used with the RT primer provided with the TaqMan® MicroRNA Assay to convert miRNA to cDNA.

A highly specific kit that quantitates only mature miRNAs, not precursors. A sensitive kit that conserves limited samples and requires only 1 to 10 nanograms of total RNA. It is fast, simple and scalable 2-step quantitative RT-PCR assay that provides high-quality results in less than three hours.

SuperScript™ IV First-Strand Synthesis System Invitrogen™

The SuperScript® IV First-Strand Synthesis System for RT-PCR is optimized for synthesis of first-strand cDNA from purified poly(A)+ or total RNA. The kit includes SuperScript® IV Reverse Transcriptase (RT), a proprietary MMLV mutant that produces robust and reliable cDNA synthesis in RT reactions. The SuperScript® IV synthesis system is significantly improved over the SuperScript® III-based system in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of the previous system, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity. The SuperScript® IV system is designed to provide reliable, consistent, and fast cDNA synthesis in the presence of inhibitors found in a wide variety of samples that cause other currently available RTs to perform inefficiently.

The SuperScript® IV First-Strand Synthesis System contains all components needed for RT reactions, plus an additional control gene and primers, and provides the flexibility to customize the RT set-up to fit the needs of your application. The SuperScript® IV synthesis system is the top choice for performance and flexibility for RT-PCR applications. We recommend it for all RT-PCR applications, especially when reproducibility and reliability is the primary concern and when inhibitors in the RNA sample may interfere with cDNA synthesis, leading to biases in gene expression studies.

Features of the SuperScript® IV First-Strand Synthesis System include:
• Significantly improved resistance to a variety of inhibitors that can interfere with cDNA synthesis
• Robust and specific cDNA synthesis in a wide range of sample types
• A faster reverse transcriptase reaction that reduces incubation time from >50 min to 10 min
• Significantly better processivity compared to SuperScript® III RT

Enzyme source
Purified from E. coli expressing the pol gene of M-MLV, modified to increase thermostability and half-life, processivity, inhibitor resistance and to reduce RNase H activity.

Performance and quality testing
Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.

Unit definition
One unit of SuperScript® IV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min at 37°C using poly(A) oligo(dT)12-18 as a template/primer.

Unit reaction conditions
50 mM Tris-HCl (pH 8.3), 4 mM MgCl2, 10 mM DTT, 50 mM KCl, 0.5 mM dTTP, 0.4 MBq/mL [3H]-dTTP, 0.4 mM poly(A) oligo (dT)12-18 and enzyme in 20 μL for 10 min at 37°C.

Gateway™ Vector Conversion System with One Shot™ ccdB Survival Cells Invitrogen™

The Gateway® Vector Conversion System is designed to convert any vector into a Gateway® destination vector using restriction endonucleases and ligase. A Gateway® cassette containing attR recombination sites flanking a ccdB gene (1) and a chloramphenicol-resistance gene are blunt-end cloned into the multiple cloning site of any vector. ccdB Survival 2 competent cells allow propagation of the Gateway® vectors containing the ccdB gene. The Gateway® Vector Conversion System offers:
• Conversion of any expression vector into a Gateway® destination vector
• Three cassettes to construct a destination vector in the correct reading frame. Each cassette has a unique restriction site to easily distinguish the cassettes and screen for the correct orientation.

Verso cDNA Synthesis Kit Thermo Scientific™

Thermo Scientific Verso cDNA synthesis Kit provides robust transcription of RNA to create a complete cDNA pool. The Verso system achieves robust and sensitive reverse transcription through the combination of a high affinity RT enzyme, a unique RNA priming method, and an optimized buffering system. The Verso RT enzyme has high RNA template affinity and reduced RNase H activity, to transcribe even sections with high secondary structure. The included anchored oligo dT priming method further enhances sensitivity by increasing transcription efficiency, and the cDNA synthesis buffer has been optimized to achieve a full and diverse cDNA pool.

Highlights

• Highly sensitive with a broad dynamic range: can reverse transcribe RNA from an input range of 1pg to 1µg
• Efficient RT reaction allows for 75% shorter protocol times (down to 30 minutes)
• Unique priming strategy for increased efficiency and broad template flexibility
• High affinity RT enzyme effectively transcribes through difficult RNA sequences
• Wide working temperature range (42°C to 57°C) for success with GC-rich and other difficult templates
• Included RT Enhancer prevents genomic DNA carryover, eliminating the need for separate DNase I treatment
• RNase Inhibitor included
• Can be paired with any PCR Kit for complete 2-Step RT-(q)PCR

Applications

• 2-step RT-(q)PCR
• Single stranded cDNA synthesis

Related Products
Verso cDNA Synthesis Kit

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).

SuperScript™ IV CellsDirect™ cDNA Synthesis Kit Invitrogen™

The SuperScript IV CellsDirect cDNA Synthesis Kit is optimized for the synthesis of first-strand cDNA directly from a mammalian cell lysate without first isolating RNA. Cell lysis and reverse transcription are performed in the same tube, and the resulting first-strand cDNA is ready to use in PCR and qPCR.

Features of the SuperScript IV kit include:
• Compatible with a wide range of mammalian cell types grown under different treatment conditions
• Single-tube format minimizes reagent loss, sample loss, and handling time
• Total lysate volume can be used in first-strand cDNA synthesis reaction, providing greater yields with a limited number of cells and allowing for detection of rare transcripts
• High-quality cDNA generated for use in a variety of applications, including PCR and qPCR
• Simple protocol takes around 37 minutes
• Compatible with TaqMan and SYBR Green qPCR master mixes from various manufacturers
• Optimized performance for 1–10,000 cells per sample, results equivalent to those from purified RNA

The SuperScript IV CellsDirect cDNA Synthesis Kit includes SuperScript IV Reverse Transcriptase. Features of this enzyme include:
• Reduced RNase H activity
• Higher thermal stability
• High yields of cDNA in the first-strand synthesis reaction, for greater sensitivity and enhanced detection of rare transcripts
• Superior tolerance to various RT inhibitors

SuperScript IV reverse transcriptase is a top choice for all RT-PCR and qRT-PCR applications. It is a proprietary MMLV mutant with superior robustness and reliability in RT reactions. It is significantly improved over SuperScript III reverse transcriptase in inhibitor resistance, processivity, and reaction speed, while retaining all the benefits of the previous enzyme, including increased thermostability, highly efficient full-length cDNA synthesis, and reduced RNase activity.

How it works
In traditional RT-PCR, RNA is first isolated from cells in a time-consuming procedure that can lead to a loss of material. Using the SuperScript IV CellsDirect cDNA Synthesis Kit, the cells are lysed and the cDNA is generated from the lysate in a single tube with minimal handling and no sample loss. DNase I treatment is an optional step and can be used to eliminate genomic DNA prior to first-strand synthesis. This kit has been optimized for small cell samples, ranging from 10,000 cells down to a single cell.

SuperScript™ III CellsDirect™ cDNA Synthesis Kit Invitrogen™

The SuperScript® III CellsDirect cDNA Synthesis Kit is optimized for synthesizing first-strand cDNA directly from a mammalian cell lysate without first isolating the RNA. Lysis and reverse transcription are performed in the same tube, and the resulting first-strand cDNA is ready to use in cloning and PCR. For real-time quantitative RT-PCR, see the note below. The kit has the following advantages:

• Compatible with a wide range of mammalian cell types grown under different treatment conditions
• Single-tube format minimizes reagent loss, sample loss, and handling time
• Total lysate volume is used in first-strand cDNA synthesis reaction, providing greater yields with a limited number of cells and allowing for detection of rare transcripts
• SuperScript® III Reverse Transcriptase, with reduced RNase H activity and higher thermal stability, produces high yields of cDNA in the first-strand synthesis reaction, for greater sensitivity and enhanced detection of rare transcripts
• Generates high-quality cDNA for use in a variety of applications, including cloning and PCR
• Simple protocol takes less than 2 hours

How it works
In traditional RT-PCR, RNA is first isolated from cells in a time-consuming procedure that can lead to a loss of material. Using the SuperScript® III CellsDirect cDNA Synthesis System, the cells are lysed and the cDNA is generated from the lysate in a single tube with minimal handling and no sample loss. DNase I is added to eliminate genomic DNA prior to first-strand synthesis. This kit has been optimized for small cell samples, ranging from 10,000 cells down to a single cell (as measured by serial dilution). The use of SuperScript® III Reverse Transcriptase ensures high specificity and high yields of cDNA from small amounts of starting material—as little as 10 pg total RNA. After synthesis, the first-strand cDNA can be amplified with specific primers by PCR without intermediate organic extractions or ethanol precipitations.

High-Capacity cDNA Reverse Transcription Kit Applied Biosystems™

The High Capacity cDNA Reverse Transcription Kit contains all components necessary for the quantitative conversion of up to 2 µg of total RNA to single-stranded cDNA in a single 20 µL reaction. The High Capacity cDNA Reverse Transcription Kit delivers extremely high-quality, single-stranded cDNA from 0.02 to 2 µg total RNA. Reactions can be scaled up to 100 µL to generate 10 µg of cDNA from a single reaction. Downstream applications include real-time PCR, standard PCR, and microarrays. The kit is ideal for generating cDNA archives.

Features of this kit include:

• Linear target amplification for real-time PCR
• Higher yields and precision than other cDNA synthesis kits at a fraction of the cost
• 10-fold greater dynamic range than other kits

Extensively tested with a variety of templates
Quantitative first-strand synthesis of all RNA species is achieved with the use of the random primers. The kit has been extensively tested with a variety of RNA templates, including GC- and AU-rich targets and RNAs expressed at low levels. cRNA can also be efficiently generated from in vitro transcription of cDNA.

Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase Thermo Scientific™

Thermo Scientific Maxima First Strand cDNA Synthesis Kit for RT-qPCR with dsDNase is a convenient system optimized for cDNA synthesis in 2-step quantitative RT-PCR (RT-qPCR) applications. It provides a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into one-tube procedure. The kit contains a novel double-strand specific DNase (dsDNase) engineered to remove contaminating genomic DNA from RNA preps in 2 minutes without damage to quality or quantity of RNA. Highly specific activity towards double-stranded DNA ensures that single-stranded DNA (such as cDNA and primers) is not cleaved and dsDNase treated RNA can be directly added to reverse transcription.

For the first strand cDNA synthesis the kit uses Maxima Reverse Transcriptase (RT), an advanced enzyme derived by in vitro evolution of M-MuLV RT. The enzyme features high thermostability, robustness and increased cDNA synthesis rate compared to wild type M-MuLV RT. The Maxima First Strand cDNA Synthesis Kit for RT-qPCR is capable of reproducible cDNA synthesis from a wide range of total RNA amounts (1 pg to 5 µg) at elevated temperatures (42 to 65°C). The synthesis reaction can be completed in 15 to 30 minutes. Components of the Maxima First Strand cDNA Synthesis Kit for RT-qPCR are pre-mixed to save time and to reduce the possibility of pipetting errors.

Highlights

• Integrated gDNA removal step
• High yields of full length cDNA up to 20 kb.
• Efficient cDNA synthesis at wide temperature range from 42°C to 65°C.
• Increased synthesis rate—complete cDNA synthesis in 15-30 minutes.
• High sensitivity and specificity.

Applications

• 2-step RT-PCR
• 2-step RT-qPCR

Maxima First Strand cDNA Synthesis Kit with dsDNase contains
• Maxima Enzyme Mix
• 5X Reaction Mix
• dsDNase
• 10X dsDNase Buffer
• nuclease-free water

Additional Information about Reaction Components
The Maxima Enzyme Mix contains Maxima Reverse Transcriptase and Thermo Scientific RiboLock RNase Inhibitor,. The recombinant RiboLock RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B and C at temperatures up to 55°C.

5X Reaction Mix contains the remaining reaction components: reaction buffer, dNTPs, oligo(dT)18 and random hexamer primers.

Water, nuclease-free is provided for reaction set-up and dilution of sample RNA. The absence of endo-, exodeoxyribonucleases, ribonucleases and phosphatases has been confirmed by appropriate quality tests.

Related Products
Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase
Maxima First Strand cDNA Synthesis Kit for RT-qPCR
Maxima First Strand cDNA Synthesis Kit Reaction Mix (5X)

NGS Reverse Transcription Kit Ion Torrent™

The Ion Torrent NGS Reverse Transcription (RT) Kit is the first cDNA synthesis kit developed specifically for next-generation sequencing (NGS) applications. It combines the superior performance of SuperScript IV Reverse Transcriptase with a novel master mix optimized for NGS library preparation. The NGS RT Kit is compatible with Ion AmpliSeq and Ion AmpliSeq HD assays and can be used with both manual and automated workflows.

Benefits of the NGS RT Kit include:
• Optimized for a variety of sample types, including FFPE, degraded samples, and those with low template amounts
• Superior NGS results with novel RT formulation optimized for NGS library preparation
• Helps increase NGS library yield, improve detection of long amplicons, and rescue under-performing amplicons
• Helps maximize mapped reads and improve detection of RNA fusions
• Trusted SuperScript IV RT technology offers superior cDNA synthesis performance with even the most challenging RNA samples
• Universal kit for wide variety of Oncomine oncology research assays and Ion AmpliSeq panels

Simple reaction setup
The NGS RT Kit contains a tube of 10X SuperScript IV enzyme mix and a tube of 5X buffer. A one-step, 25-minute reaction reverse transcribes total RNA into cDNA and can be used directly for NGS library preparation, without any additional quantification, purification, or dilution steps. A variety of sample types can be used, including RNA from blood, bone marrow, FFPE and partially degraded samples. This product is compatible with Oncomine and Ion AmpliSeq workflows, as well as Ion AmpliSeq HD panels. Consult the cDNA synthesis section of each assay manual for specific RT reaction volumes and setup instructions.

Performance and quality testing
The NGS RT Kit is assayed for endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease, as well as yield and length of cDNA product. It is QC tested with a full NGS workflow to evaluate library yield and ensure performance meets key sequencing metrics.

GeneArt™ Gibson Assembly EX Master Mix Invitrogen™

GeneArt Gibson Assembly EX Master Mix enables DNA assembly via a technique that allows multiple overlapping DNA fragments to be seamlessly linked in a single-tube, two-step optimized reaction. DNA fragments of different lengths are uniformly assembled using complementary overlaps between fragments. The inherent flexibility and success of this approach is suitable for small and large DNA constructs, includes both single and multiple inserts, and has been used to build entire genomes. The resulting products can be used for a variety of downstream applications, including transformation, PCR, and rolling circle amplification (RCA). The GeneArt Gibson Assembly EX Master Mix kit includes master mix, positive control, and water, and accommodates the use of your own competent cells.

Features of the GeneArt Gibson Assembly EX Master Mix include:
Simple—seamlessly assemble and clone up to 15 DNA fragments in a single reaction
Efficient—robust efficiency provides successful clones for both simple and challenging constructs
Flexible—design guidelines allow assembly into any vector of your choice
Convenient—available as master mix kits and cloning kits complete with chemical or electrocompetent cells
Trusted—over 4000 citations in the scientific literature highlighting great success

Seamless assembly of up to 15 fragments
GeneArt Gibson Assembly EX cloning is a robust, single-tube, two-step process whereby up to 15 inserts and vector are combined in a proprietary enzymatic mix in order to assemble DNA fragments with shared terminal end homology without leaving any extra sequences or scars behind (seamless). After this dually optimized reaction is complete, a portion of the assembly reaction is then transformed into chemically competent or electrocompetent cells, yielding clones ready for analysis the next day. The required 20- to 40-base pair end homology is designed into the de novo fragment for synthesis or can be easily engineered by PCR amplification with custom DNA oligos. This special enzyme mix creates a seamless and covalently bound DNA construct providing high efficiency.

Robust method for maximum efficiency
Due to the covalently bound final product, the GeneArt Gibson Assembly EX method allows the utilization of chemically competent cells or electrocompetent cells for the highest transformation efficiency, especially important for challenging constructs. There is no need for restriction enzymes, ligation, or recombination sites, and the method provides a perfectly seamless construct without unwanted extra bases.

Provides great versatility
The GeneArt Gibson Assembly EX method provides versatility, can streamline many techniques through the rapid combination, addition, deletion, or exchange of DNA segments, and eliminates the need to subclone, saving time and effort in the cloning workflow.

Convenient formats
GeneArt Gibson Assembly EX kits are available in multiple formats, including:
GeneArt Gibson Assembly EX Cloning Kit, electrocompetent cells (with ElectroMAX DH10B electrocompetent cells)
GeneArt Gibson Assembly EX Cloning Kit, chemically competent cells (with One Shot TOP10 chemically competent cells)
• GeneArt Gibson Assembly EX Master Mix kits (cloning kit without the cells) (this page)

SuperScript™ VILO™ cDNA Synthesis Kit Invitrogen™

The SuperScript VILO cDNA Synthesis Kit is designed to generate first strand cDNA for two-step RT-qPCR applications. The kit is supplied in a 2-tube format with the VILO Reaction Mix and an enzyme blend in separate tubes. The enzyme blend contains SuperScript III Reverse Transcriptase (RT), a genetically engineered MMLV RT that has reduced RNase H activity and improved thermostability for highly efficient cDNA synthesis. The kit can be used to synthesize cDNA from for a wide range of input RNA amounts.

Note: For superior first-strand cDNA synthesis performance in two-step RT-qPCR, we recommend SuperScript IV VILO Master Mix or SuperScript IV VILO Master Mix with ezDNase Enzyme. The master mix provides all cDNA synthesis reaction components in a convenient one-tube master mix format. It elevates the trusted VILO technology to the next level with the highly processive and thermostable SuperScript IV RT and further optimized buffer. The master mix enables efficient cDNA synthesis at higher temperatures in less time and provides superior cDNA yield and sensitivity even with suboptimal purity or scarce templates.

GeneArt™ Seamless PLUS Cloning and Assembly Kit Invitrogen™

Like our first generation Seamless Cloning and Assembly Kit, GeneArt® Seamless PLUS Cloning and Assembly Kit is the complete kit for simultaneous and directional cloning of 1 to 4 PCR fragments, consisting of any sequence, into any linearized vector, in a single 30-minute or less, room temperature reaction. However, Seamless PLUS offers several advantages over previous kits:

Increased Efficiency: pre-cloning option for large fragments for increased cloning efficiency
Larger Constructs: create constructs up to 40 kb
Versatility: high capacity, broad-range conjugative vector that replicates in most Gram negative bacteria

The improvements above are combined with these key benefits shared by all GeneArt® Seamless Cloning and Assembly kits:

Speed and Ease—clone up to 4 DNA fragments, with sequence of your choice, simultaneously in a single vector; no restriction digestion, ligation, or recombination sites required
Precision and Efficiency—designed to let you clone what you want, where you want, in the orientation you want, and achieve up to 90% correct clones with no extra sequences left behind
Free Tools—design your final construct and DNA oligos in silico using our free web-based tool that takes you step-by-step through your project
• Vector Flexibility
—use our linear vector or a vector of your choice
Diverse Applications—streamline many synthetic biology and molecular biology techniques through the rapid combination, addition, deletion, or exchange of DNA segments

For cloning of more than 4 DNA fragments or for final molecules larger than 110 Kb, please consider the GeneArt® High-Order Genetic Assembly System.

Simple and Fast Clone Creation
GeneArt® Seamless PLUS Cloning is a simple, two-step process, consisting of in vitro assembly followed by transformation into One Shot® DH10B™ T1R SA competent E. coli. The kit employs a proprietary enzyme/buffer mix to assemble DNA fragments with shared terminal end homology without extra sequences or scars in the final construct ('seamless'). Terminal end homology is easily incorporated by PCR amplification with custom DNA oligos engineered using our free web tool.

Cloning Efficiency, Flexibility, and Precision
With the GeneArt® Seamless PLUS Cloning and Assembly Kit, the main factors effecting cloning efficiency are the size of the DNA fragments (100 bp to 10 Kb), the total size of the final molecule (≤ 40 Kb), and the quality and specificity of each fragment.

Typical cloning efficiencies for different numbers of fragments:

• >95% for 4 fragments, 5 Kb each
• >90% for 4 fragments, 10 Kb each

Cloning success is independent of the insert sequence and vector type, allowing you to design and add nearly any desired sequence, or combination of sequences, to any plasmid as long as it can be linearized by either restriction enzyme digestion or PCR. The circularized clones obtained from the reaction contain only the sequence of your original vector, inserts, and designated homologies, with no extraneous nucleotide insertions.

in silico Design Support
A key step in GeneArt® Seamless PLUS Cloning is the correct design of fragments and oligos with the appropriate homology and spacing to help ensure successful assembly of your clone. We provide a free online tool, the GeneArt® Design Tool for Seamless or High-Order Assembly and Mutagenesis, to help you design your experiment in silico. The tool checks for compatibility of the experimental design with the product specifications, designs DNA oligos with end homology for the PCR amplification of the different elements to clone, and presents the user with a graphical representation of the vector, as well as a downloadable annotated sequence in GenBank format that is compatible with Vector NTI® software.

Applications
The GeneArt® Seamless PLUS Cloning and Assembly Kit is designed to empower cloning and DNA assembly in a wide range of molecular biology and synthetic biology applications, among others. The product allows for the creation of modular expression vectors, with interchangeable parts, and can be used to perform a variety of tasks that would otherwise involve multiple steps. Use the kit to construct fusion proteins; delete, replace, or add DNA elements such as restriction sites in an existing vector; and carry out many other techniques that require manipulation of genetic sequences.

Maxima First Strand cDNA Synthesis Kit Reaction Mix (5X) Thermo Scientific™

Thermo Scientific Maxima 5X Reaction Mix is a component of the Maxima First Strand cDNA Synthesis Kits for RT-qPCR (K1671/K1672/K1641/K1642) and may be purchased separately. The 5X Reaction Mix is comprised of reaction buffer, dNTPs, oligo (dT)18 and random hexamer primers.

Related Products
Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase
Maxima First Strand cDNA Synthesis Kit for RT-qPCR
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