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SuperScript™ III First-Strand Synthesis SuperMix for qRT-PCR (Invitrogen™)

SuperScript® III First-Strand Synthesis SuperMix for qRT-PCR provides the high-temperature capability of SuperScript® III Reverse Transcriptase in an optimized SuperMix format for the synthesis of first-strand cDNA for use in real-time quantitative RT-PCR (qRT-PCR). The simple, time-saving reaction set-up uses just two tubes: a 2X Reaction Mix and an Enzyme Mix.

Enzyme mix
SuperScript® III Reverse Transcriptase, included in the RT Enzyme Mix, is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can be used to synthesize cDNA at a temperature range of 42–60°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 can be used to synthesize cDNA from total RNA. RNaseOUT™ Recombinant Ribonuclease Inhibitor, also included in the enzyme mix, is an RNase inhibitor protein that safeguards against the degradation of target RNA due to ribonuclease contamination of the RNA preparation.

Reaction mix
The 2X RT Reaction Mix includes oligo(dT)20, random hexamers, MgCl2, and dNTPs in a buffer formulation that has been optimized for qRT-PCR. E. coli RNase H is provided as a separate tube in the kit to remove the RNA template from the cDNA:RNA hybrid molecule after first-strand synthesis. This has been shown to increase sensitivity in qRT-PCR.

Using SuperScript® III First-Strand Synthesis SuperMix
This SuperMix formulation can be used to quantify fewer than 10 copies of a target gene in qRT-PCR, with a broad dynamic range that supports accurate quantification of high-copy mRNA from up to 1 µg of total RNA. Reagents are provided for 50 or 250 RT reactions of 20 µL each.

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

RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific™)

Thermo Scientific RevertAid First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. The kit uses RevertAid Reverse Transcriptase (RT), a recombinant M-MuLV RT which maintains activity at 42-50°C and is suitable for synthesis of cDNA up to 13 kb. RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA templates from degradation.
The RevertAid First Strand cDNA Synthesis Kit is supplied with both oligo(dT)18 and random hexamer primers. The oligo(dT)18 primer anneals selectively to the poly(A) tail of mRNA. Random hexamer primers do not require the presence of the poly(A) tail, therefore, they can be used for transcription of the 5'-end regions of mRNA or cDNA synthesis of RNA species lacking a poly(A) tail (e.g., microRNAs). Gene-specific primers may also be used with the kit.

Highlights

• Full-length first strand cDNA up to 13 kb
• Optimum reaction temperature 42°C
• Complete kit—all the components for the RT reaction are included

Applications

• First strand cDNA synthesis for RT-PCR and RT-qPCR
• Construction of full length cDNA libraries
• Antisense RNA synthesis

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.

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.

Maxima™ H Minus cDNA Synthesis Master Mix (Thermo Scientific™)

Thermo Scientific Maxima H Minus cDNA Synthesis Master Mix provides all cDNA synthesis reaction components in a convenient one-tube master mix. It is optimized for highly efficient cDNA synthesis for two-step quantitative RT-PCR (RT-qPCR) applications.

Features include:
• One-tube master mix helps reduce pipetting steps and enhanced consistency in RT-qPCR results
• Increased RT efficiency across a wide range of input RNA amounts and gene targets
• High thermostability to allow RT reactions at 50 to 65°C temperature range

The master mix contains Maxima H Minus Reverse Transcriptase (RT) and RiboLock RNase Inhibitor. Maxima H Minus RT is an advanced enzyme derived from M-MuLV RT by in vitro evolution. The enzyme features high thermostability, robustness, processivity, increased cDNA synthesis rate, and lack of RNase H activity. The recombinant RiboLock RNase Inhibitor effectively protects RNA template from degradation by RNases A, B, and C at temperatures up to 55°C. The master mix also contains reaction buffer, dNTPs, oligo (dT)18, and random hexamer primers. A separate tube of No-RT control mix is also provided.

Maxima H Minus cDNA Synthesis Master Mix is capable of reproducible cDNA synthesis at elevated temperatures (50–65°C). The synthesis reaction is typically complete in 15–30 minutes.

Additional information about reaction components
• The No RT Control mix contains all the components in the Maxima H Minus cDNA Synthesis Master Mix except RT. The presence of a PCR product in the No RT Control reaction indicates that the reaction is contaminated with DNA. To further enhance genomic DNA elimination efficiency, template RNA incubation with dsDNase (Cat. No.EN0771) is strongly recommended.
• Nuclease-free water is provided for reaction set-up and dilution of sample DNA. The absence of exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.

Related products
Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase
Maxima H Minus First Strand cDNA Synthesis Kit
Maxima H Minus Reverse Transcriptase
dsDNase

Arcturus™ RiboAmp™ HS PLUS cDNA Kit (Applied Biosystems™)

The ARCTURUS® RiboAmp® HS PLUS cDNA Kit permits quantitative Real-Time PCR (qRT-PCR) from as little as 100pg of total RNA. The kit uses random primers to enable robust reverse transcription across a broader transcript length.

Integrated Systems for Microgenomics
The ARCTURUS® Complete System for Microgenomics® offers an integrated solution for preparing small quantities of RNA for gene expression analysis. The System features the ARCTURUSXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations, the PicoPure® RNA Isolation Kit to extract and purify RNA, and the RiboAmp® PLUS Kit for reverse transcription and linear amplification of RNA. ARCTURUS® Systems for Microgenomics enable accurate and sensitive qRT-PCR and microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

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

RevertAid H Minus First Strand cDNA Synthesis Kit (Thermo Scientific™)

Thermo Scientific RevertAid H Minus First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. The kit includes RevertAid H Minus Reverse Transcriptase, which has a point mutation that completely eliminates RNase H activity. Therefore, it does not degrade RNA in RNA-DNA hybrids during synthesis of the first strand cDNA.

Highlights

• High yields of full-length first strand cDNA up to 13 kb
• Increased reaction temperatures in the range of 42 to 55°C
• Supplied with the recombinant RiboLock RNase Inhibitor
• Complete kit—oligo(dT)18 and random hexamer primers included with the kit

Applications

• First strand cDNA synthesis for RT-PCR and real-time RT-PCR
• Construction of full length cDNA libraries
• Antisense RNA synthesis

Additional Features
The recombinant RiboLock RNase Inhibitor, supplied with the kit, effectively protects RNA template from degradation. It is fully compatible with the reverse transcription reaction, as it maintains activity at temperatures up to 55°C.The kit is supplied with both oligo(dT)18 and random hexamer primers. The oligo(dT)18 primer anneals selectively to the poly(A) tail of mRNA. Random hexamer primers do not require the presence of poly(A), therefore, they can be used for transcription of the 5’-end regions of mRNA or for cDNA synthesis using RNA without the poly(A) tail, e.g., microRNAs. Gene-specific primers may also be used with the kit. The first strand cDNA can be directly used as a template in PCR, realtime PCR or in second strand cDNA synthesis.

GeneArt™ Site-Directed Mutagenesis PLUS System (Invitrogen™)

The GeneArt® Site-Directed Mutagenesis PLUS System brings our mutagenesis technology to the next level with the ability to perform single- or multi-site mutations in larger plasmids with greater efficiency and in less time than the competition.

Powerful—make substitutions, deletions, or insertions of up to 3 nucleotides in 1, 2, or 3 separate sites in DNA plasmids of up to 14 kb
Flexible—perform multi-site mutagenesis with degenerated nucleotides and single-site mutations up to 25 nucleotides long or 12 nucleotides long with degenerated nucleotides
Efficient—obtain your desired mutants the first time; over 90% correct mutants in a 10 Kb plasmid
Fast—obtain your mutated plasmid DNA in typically less than 3 hours with the simple, minimal-handling protocol
Versatile—use plasmids of many sizes, and DNA isolated from any source, with no need for specialized vectors, host strains, or restriction sites

Optimized Mutagenesis Protocol
The GeneArt® Site-Directed Mutagenesis System was further optimized for efficiency and multi-site capability, resulting in the "PLUS" kit. Like the first generation GeneArt® Site-Directed Mutagenesis System, DNA methylation and amplification steps are combined into a single reaction, with no need for an in vitro DpnI digestion step. After methylation and amplification, a 15 minute in vitro recombination reaction of the amplified PCR products increases the colony output by 3 to 10 fold, resulting in a higher mutagenesis efficiency. A final transformation of the mutated DNA into DH5α™ E.coli cells digests any methylated parental DNA, leaving behind only the intact unmethylated mutagenesis reaction product. No purification steps or additional digestions are needed. Individual colonies can be selected the following day to verify mutations.

Simple Creation of Desired Mutants
Creating mutants with the GeneArt® Site-Directed Mutagenesis PLUS System relies on the inherent properties of DNA methylase, high fidelity DNA polymerase, recombination enzymes, and the native McrBC endonuclease of E. coli. Simply incorporate your desired mutation or mutations into primers using our GeneArt® Design Tool for Seamless Assembly and Mutagenesis (see "in silico Design Support", below). Combine the vector and mutagenesis primers, and after PCR, recombination, and transformation you will have vectors with only the desired mutations with over 90% efficiency.

in silico Design Support
A key step in GeneArt® site-directed mutagenesis is the correct design of primers with the appropriate homology and spacing to help ensure successful mutagenesis of your plasmid. To simplify and speed the design process, 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 allows you to review your plasmid sequence, designs DNA oligos for introduction of the desired mutations, provides an updated amino acid sequence for your mutated gene, displays a graphical representation of the new construct, and enables the download of an annotated sequence in GenBank format that is compatible with Vector NTI® software.

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

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.

Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase (Thermo Scientific™)

Thermo Scientific Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase 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 dsDNase 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 reverse transcription the kit uses Thermo Scientific Maxima H Minus Reverse Transcriptase (RT), which is an advanced enzyme derived by in vitro evolution of M-MuLV RT. The enzyme features the highest thermostability among the derivatives of M-MuLV RT and lacks RNase H activity. The Maxima H Minus First Strand cDNA Synthesis Kit allows synthesis of long cDNAs up to 20 kb at elevated temperatures (up to 65°C), superseding other systems' abilities to produce full-length cDNA. Due to increased synthesis rates the reaction can be completed in 30 minutes.

Highlights

• Integrated genomic DNA removal step
• Increased reaction temperatures—the first strand of cDNA can be synthesized within the 42 to 65°C temperature range
• High yields of full-length first strand cDNA—with RNA templates up to 20 kb
• Flexible priming—oligo(dT)18, random hexamer, or gene-specific primers

Applications

• First Strand cDNA synthesis for RT-PCR
• Construction of cDNA libraries
• Generation of probes for hybridization
• Antisense RNA synthesis

Includes

Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase contains Maxima H Minus Enzyme Mix, dsDNase, 10X dsDNase Buffer, Oligo(dT)18 and Random hexamer primers, 5X RT Buffer, dNTP Mix, and nuclease-free water.

Additional information about reaction components

• Maxima H Minus Enzyme Mix contains Maxima H Minus Reverse Transcriptase and RiboLock RNase Inhibitor. RiboLock RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B and C at temperatures up to 55°C.
oligo(dT)18 and random hexamer primers are supplied with the kit. Random hexamer primers bind non-specifically and are used to synthesize cDNA from all RNAs in a total RNA population. The oligo(dT)18 primer selectively anneals to the 3'-end of poly(A) RNA, synthesizing cDNA only from poly(A) tailed mRNA. Gene-specific primers may also be used with the kit to prime synthesis from a specified sequence.
10 mM dNTP Mix is a premixed aqueous solution of dATP, dTTP, dCTP, and dGTP.
Nuclease-free water is provided for reaction set-up and dilution of sample DNA. The absence of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.

Related Products
Maxima H Minus First Strand cDNA Synthesis Kit, with dsDNase
Maxima H Minus First Strand cDNA Synthesis Kit

SuperScript™ III First-Strand Synthesis SuperMix (Invitrogen™)

The SuperScript® III First-Strand Synthesis SuperMix is an optimized SuperMix formulation for first-strand cDNA synthesis 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 0.1 pg to 5 µg of total RNA.

Using the SuperScript® III First-Strand Synthesis SuperMix
The kit includes SuperScript® III/RNaseOUT™ Enzyme Mix, 2X First-Strand Reaction Mix, and Annealing Buffer. SuperScript® III Reverse Transcriptase, included in the Enzyme Mix, is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can be used to synthesize cDNA at a temperature range of 45–55°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 can be used to synthesize cDNA from total RNA. RNaseOUT™ Recombinant RNase Inhibitor is included in the Enzyme Mix to safeguard against degradation of target RNA due to ribonuclease contamination. The 2X First-Strand Reaction Mix includes 10 mM MgCl2 and 1 mM of each dNTP in a buffer formulation that has been optimized for first-strand synthesis of cDNA. The Annealing Buffer is used in the initial template-primer annealing step. Separate tubes of oligo(dT)20 and random hexamers are also provided. cDNA synthesis can be performed using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer.

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

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)

GeneArt™ Seamless Cloning and Assembly Enzyme Mix (Invitrogen™)

GeneArt® Seamless Cloning and Assembly Enzyme Mix enables the simultaneous and directional cloning of 1 to 4 PCR fragments, consisting of any sequence, into any linearized vector, in a single 30-minute room temperature reaction. GeneArt® Seamless enzyme mix is the economical choice for creating constructs up to 13 kb with the option for high-throughput assembly.

Efficiency: pre-cloning option for large fragments for increased cloning efficiency
Speed and Ease: clone DNA fragments, with sequence of your choice, in a single vector (up to 13 kb); no restriction digestion, ligation, or recombination sites required
Free Tools: design of 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

For the cloning of pre-existing DNA elements too long to be amplified by PCR and for final molecules up to 40 Kb, please consider the GeneArt® Seamless PLUS Cloning and Assembly Kit (1–4 DNA fragments only); and for final molecules larger than 110 Kb, please consider the GeneArt® High-Order Genetic Assembly System.

Simple and Fast Clone Creation
GeneArt® Seamless Cloning is a simple, two step process, consisting of in vitro assembly followed by transformation. The enzyme mix uses a proprietary enzyme/buffer mix to assemble DNA fragments without extra sequences or scars ("seamless"). Terminal end homology, if needed, is easily incorporated by PCR amplification with custom DNA oligos engineered using our free web tool.

Cloning Efficiency, Flexibility, and Precision
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 Cloning Design Support
A key step in GeneArt® Seamless Cloning is the correct design of fragments and oligos with the appropriate homology and spacing to help ensure successful assembly of your clone. To simplify and speed the design process, 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 guides the user through construct design, including fragment import, order, and orientation; checks for areas of homology and potential design issues; creates primers for pre-cloning or incorporating end homology between fragments, if needed; and generates final construct map files and sequence in Genbank format for download or import into Vector NTI® software.

Applications
The GeneArt® Seamless Cloning and Assembly Enzyme Mix 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 mix 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.

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