Invitrogen cloning vectors.
The TA Cloning Kit with pCR 2.
Invitrogen cloning vectors Before cloning your own reagent, you might be able to save time by finding it in one of the many well-characterized collections. K4120-01 and K5120-01) available from Invitrogen. The TA Cloning Kit with pCR 2. 7k Invitrogen Corporation 1600 Faraday Avenue Carlsbad, CA 92008 T: 1 760 603 7200 F: 1 760 602 6500 E: tech. Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene Try SnapGene for Free Download Plasmid | Explore Over 2. Other reading frames are available with pENTR 2B and pENTR 3C. com pBAD/His A, B, and C pBAD/Myc-His A, B To further simplify and streamline the cloning workflow, specialized vectors have been developed to place an insert into vector, for example, without using a ligase. 点击了解Invitrogen Gateway重组克隆技术如何通过一步克隆，BR和LR Clonase反应实验步骤，节省基因片段组合时间，提升克隆效率。高效便捷的克隆技术 Invitrogen Gateway 重组克隆技术避免了基于限制性内切酶的传统克隆技术的各种限制因素，通过简单几步即可穿梭进几乎所有表达系统 Save up to five days of turnaround time with direct cloning into one of our select Invitrogen vectors, such as pcDNA 3. For rapid TOPO® Cloning of PCR products, we recommend using the pENTR/D Choose one of the InvitrogenTM TOPOTM cloning vectors to create your Entry clone, or custom fragment in the desired vector through. pCR -TOPO Vectors include: • 3´-T To further simplify and streamline the cloning workflow, specialized vectors have been developed to place an insert into vector, for example, without using a ligase. coli Bacterial directional TOPO® cloning vector, for inducible expression of a C-terminally V5- and 6xHis-tagged protein. The pDONR 221 vector has a pUC Traditional cloning relies on recombinant DNA methods that begin with preparing a vector to receive an insert DNA by digesting each with restriction enzymes. One such class of vectors includes the Invitrogen TOPO cloning vectors which Save up to five days of turnaround time with direct cloning into one of our select Invitrogen vectors, such as pcDNA 3. Quick—avoid inefficient ligation and laborious searches for appropriate restriction enzymes. 1 and pCR II vectors. These products include kits for cloning, transfection, and expression of DNA. Eac Quick and efficient PCR cloning with TOPO TA cloning Clone PCR-amplified DNA fragments (blunt or A-overhang) directly into a choice of over 40 subcloning, sequencing, or expression vectors in just 5 minutes—and obtain up to 95% recombinant clones. Cloning Kits Five-minute, directional TOPO® Cloning of blunt-end PCR products into an entry vector for the Gateway® System Catalog numbers K2400-20, K2420-20, K2525-20, K2535-20, K2435-20, and K2635-20 Revision Date 29 March 2012 Publication part Our CT GFP-fusion TOPO® Expression kit provides pcDNA 3. Each kit Use the following outline to clone and express your gene of interest in pcDNA 3. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular These single-nucleotide overhangs anneal to the 3´‑thymidine overhangs of T-cloning vectors such as the Promega pGEM®-T (Cat. 1. 1 cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. A choice of pENTRTM vectors is available (see Gateway® Dual Selection Vector for creating an entry vector by restriction cloning. 1-TOPO® but with an SP6 promoter. The pcDNA™3. Eac Methods Overview Introduction pMT/BiP/V5-His is a 3. The cloning vector, pZErO®ccd cloning, long-fragment cloning, expression-vector cloning, or directional cloning, or you’re using the Invitrogen Gateway system, there’s a TOPO cloning solution that’s right for you. pCR-TOPO TOPO™ TA Cloning™ Kit, with pCR™2. Similar to pCR®2. 2. 0 µl TE Buffer, pH 8. Gateway® donor vector with attP1 and attP2 sites and a kanamycin resistance marker. service@invitrogen. TOPO PCR cloning kits help you get the right clone sooner, freeing up your It contains a Geneticin selectable marker and a forward-orientation multiple cloning site. One such class of vectors includes the Invitrogen TOPO cloning vectors which contain covalently linked DNA topoisomerase I that functions as both a restriction enzyme and a ligase (learn more about TOPO cloning Bacterial directional TOPO® cloning vector with an ampicillin resistance marker, for inducible expression of an N-terminally 6xHis-tagged protein. TA Cloning ® Kit Catalog Numbers K2000-01, K2000-40, K2020-20, K2020-40, K2030-01 K2030-40, K2040-01, and K2040-40 2. The tool allows you to quickly identify a vector that best suits your needs. 1-TOPO TA v To further simplify and streamline the cloning workflow, specialized vectors have been developed to place an insert into vector, for example, without using a ligase. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular biology Zero Blunt TOPO PCR Cloning kits for subcloning provide a highly efficient, 5-minute cloning reaction for the direct insertion of blunt-ended PCR products amplified with proofreading thermostable polymerases into a plasmid vector. We offer a range of popular expression vectors for a variety of applications. It has Experience Gateway cloning technology. The Zero Blunt PCR Cloning Kit uses the multipurpose cloning vector pCR -Blunt and ExpressLink T4 DNA Ligase to generate a ligation product in a five-minute, room-temperature ligation step. Other reading frames are available with pENTR™2B and pENTR™3C. They use a pCR -TOPO Vector with covalently bound topoisomerase I for fast cloning and recombinants. 1 vector as loss of the 3´ T-overhangs will cause a blunt-end self-ligation of the vector and subsequent decrease in ligation Use the TOPO TA Cloning Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes (1). Features of the Zero Blunt™ Cloning™ Kit The Invitrogen Gateway Cloning Technology provides a rapid and highly efficient route to protein expression, functional analysis, and cloning/subcloning of DNA segments. 1 Expression Vector Family Three untagged versions of pcDNA™3. Such overhangs at the vector cloning site not only facilitate cloning, but also prevent the recircularization of the vector. 0 add to a final volume of 15 µl Converting your favorite set of cloning vectors to Gateway Technology is a fairly straightforward The TOPO TA Cloning Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO Cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. Get your de novo gene cloned into one of direct expression-ready vectors using GeneArt Instant Designer This clone was then used in an LR recombination reaction between the entry clone and the Amp-resistant destination vector pDEST15 (Invitrogen) or the Kan-resistant destination vector pMDC32 [3]. Eac Enter the Gateway platform via Invitrogen TOPO cloning vectors containing Gateway att sites or purchase an Thermo Scientific Ultimate ORF Clone already inserted into a Gateway vector New advancements such as MultiSite Gateway Technology make Gateway cloning the ideal cloning method for protein expression and functional analysis. The Invitrogen Gateway Cloning Technology provides a rapid and highly efficient route to protein expression, functional analysis, and cloning/subcloning of DNA segments. coli Catalog Numbers 11824-026, 11801-016, 11802-014, 11803-012 Gateway® Cloning is a universal cloning technique developed by Invitrogen life technologies. coli InvivoGen provides cloning and expression vectors/plasmids for RNAi, antibody production, transgenesis and mammalian gene expression. The pENTRTM vectors allow restriction cloning of a gene of interest into a vector for entry into the Gateway® System available from Invitrogen. Today, Invitrogen TOPO cloning kits are still the fastest, simplest, and most efficient cloning solutions available. It has effectively replaced the use Proporciona una estrategia de clonación de gran eficacia, de 5 minutos y de un solo paso (<i>TOPO Cloning</i>) para la inserción directa de productos PCR amplificados con polimerasa <i>Taq</i> en un vector plasmídico para subclonación. 5 ml 2 µg pYD1 Forward sequencing primer, lyophilized 2 µg pYD1 Reverse sequencing primer, lyophilized Shipping/Storage The pYD1 Vector Kit is shipped on dry ice. sites of the donor vector. Each kit uses the pCR2. Optimize discovery : Utilize GeneOptimizer tool to improve protein expression and translational efficiency for To further simplify and streamline the cloning workflow, specialized vectors have been developed to place an insert into vector, for example, without using a ligase. What links here Related changes Upload file Permanent link Page information Cite this page Get shortened URL Download QR code The Gateway cloning method is a method of molecular cloning invented and commercialized by Invitrogen since the late 1990s, which makes use of the integration and excision recombination reactions that take place when bacteriophage lambda Mammalian expression vector with the CMV promoter and a neomycin-resistance marker. This kit is designed to clone cohesive or blunt-ended DNA fragments with a low background of non-recombinants. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures Fast accurate construct design for all major molecular Tools for DNA cloning A range of products from restriction enzymes to custom gene synthesis We’ve provided high-quality tools for DNA cloning for 30 years, continually improving existing technologies and developing new ones. Explore the Scientific R&D Platform Scientific intelligence platform for AI-powered data management and workflow automation Parental vector for TOPO® cloning of blunt PCR products. ) has developed a fast and reliable alternative cloning methodology which uses Invitrogen Gateway cloning technology has been cited by life science researchers more than 2,000 times. 1 (available separately), each with a different selectable marker v Important Information Contents 20 µg of pYD1, lyophilized in TE, pH 8 EBY100 glycerol stock, 0. The technology described here is based on vectors containing the lethal E. Simply filter by preferred Twenty years ago, the TOPO name became synonymous with superior quality and game-changing innovation in PCR cloning. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures Fast accurate construct vector (e. InvitrogenTM GeneArtTM services. 1 provides a quick, one-step cloning strategy for the directly inserting a PCR product into a plasmid vector. 3. com For country-specific contact information visit our web site at www. Generate the desired expression clone by performing an LR recombination reaction between the entry clone and a Gateway ® destination vector of choice. Our pCR8TOPO TA vector contains a minimal MCS so you sequence more of yourins Gateway Cloning: Top 5 Tips 1. coli gene, ccdB (Bernard, et al. Upon transfection, the vector allows Invitrogen GeneArt Seamless Cloning and Assembly Kits enable in vitro cloning of up to 4 DNA fragments simultaneously into virtually any linearized vector, typically in 30 minutes, without extra DNA sequences, restriction endonucleases, or ligation. Methods Overview Introduction The pENTR Dual Selection Vectors allow restriction cloning of a gene of interest into a vector for entry into the Gateway® System available from Invitrogen. A choice of pENTR Dual Selection Vectors is available (see table below cloning, long-fragment cloning, expression-vector cloning, or directional cloning, or you’re using the Invitrogen Gateway system, there’s a TOPO cloning solution that’s right for you. The pYD1 Vector Kit is shipped on dry ice. pENTR vector). These single-nucleotide overhangs anneal to the 3´‑thymidine overhangs of T-cloning vectors such as the Promega pGEM®-T (Cat. Simply filter by preferred features, including host system, selection marker, promoter, or other key characteristics. pDONR 221) and an att B PCR product or expression clone. 1⁄CT-GFP-TOPO® vector for highly efficient, 5-minute cloning that fuses the green fluorescent protein (GFP) to the C-terminal end your Taq polymerase-amplified gene product. competent 25 TOPO ® TA Cloning ® Kit Dual Promoter (with pCR II-TOPO ® vector) Background In plant functional genomic studies, gene cloning into binary vectors for plant transformation is a routine procedure. pCR4-TOPO allows you to directly select Invitrogen GeneArt Strings DNA Fragments We offer custom-made, uncloned, linear DNA fragments from 200 to 3,000 bp in length—ready for cloning and screening with any vector of Gateway® Cloning is a universal cloning technique developed by Invitrogen life technologies. USER GUIDE E. One such class of vectors includes the Invitrogen TOPO cloning vectors which contain covalently linked DNA topoisomerase I that functions as both a restriction enzyme and a ligase (learn more about TOPO cloning Cloning efficiencies are exceptionally high, regard-less of PCR product size, and yield minimal back-ground (Table 2). # A3600) and pGEM®-T Easy Vectors (Cat. The kits include linearized and topoisomerase I Use the following outline to clone and express your gene of interest in pcDNA 3. www. Use SnapGene to verify The TOPO TA Cloning Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO Cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. Ligate your insert into the appropriate vector andE. In recent years, however, Gateway® cloning technology (Invitrogen Co. The TA Cloning™ Kit uses the pCR™2. Jump to navigation Menu 888 457 5873 Contact us Need a quote ? USA USA Quick Order Add Login Cart (0) Products The recent development of transformation vectors and large-scale clone resources amply demonstrate that plant researchers have adopted the Gateway platform and that it will remain an important asset in projects requiring systematic cloning, modular assembly The TOPO TA Cloning Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy (TOPO Cloning) for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. coli Expression System with Gateway ® Technology Gateway ®-adapted destination vectors for cloning and high-level expression of native or tagged recombinant proteins in E. 3 µl BP Clonase II enzyme mix 3. TOPO PCR cloning kits help you get the right clone sooner, freeing up your TOPO TA Cloning kits for subcloning provide a highly efficient, 5-minute cloning reaction for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for subcloning. The TOPO TA Cloning Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO Cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. TOPO PCR cloning kits help you get the right clone sooner, freeing up your attP DNA (Invitrogen pDONR vector, 150 ng/µl) 1. Gateway technology relies on the two reactions described below: The BP Reaction takes place between the att B sites flanking the insert and the att P sites of the donor vector. Buy it Gateway is a high throughput method for cloning, vis a vis, it is FAST. 1 vector An Entry clone contains your gene of interest flanked by attL sequences, which are then used to recombine with attR sequences to create your desired expression clone. Introduce your 5 Figure 1. It’s no wonder Gateway cloning has been the go-to choice for years, by researchers with varying experience—from beginners to advanced—for protein Once you have an entry clone, you can recombine your gene of interest into a variety of expression vectors adapted for use in Gateway Technology. , 1994). Explore the full spectrum of Invitrogen products for molecular cloning workflow: vectors, gene synthesis and assembly tools, and other essentials. 这样一类载体包括 Invitrogen TOPO 克隆载体，其含有共价连接的DNA拓扑异构酶I，可同时作为限制性内切酶和连接酶（详细了解 TOPO cloning technology）。 与传统 PCR 克隆载体相比，这类载体具有更短的连接反应时间（如，5 分钟）、更高的克隆效率（如，>95％ 的阳性克隆）以及更简单的分子克隆实验方案。 PCR amplification or restriction-enzyme cloning vectors pDONR and pENTR vectors These vectors allow you to clone a PCR product amplified with primers containing attB sequences (Invitrogen pDONR vector) or specific restriction sites (Invitrogen pENTR vector). As a result, more than 90% of recombinant Flexible, custom process: Obtain the exact sequence you need in the vector of choice—including challenging, complex constructs and custom vectors. One such class of vectors includes the Invitrogen TOPO cloning vectors which contain covalently linked DNA topoisomerase I that functions as both a restriction enzyme and a ligase (learn more about TOPO cloning Invitrogen TOPO TA Cloning Kits for Sequencing utlize our popular Invitrogen pCR4-TOPO TA Vector for the direct ligation of Taq-amplified PCR products in a 5-minute, benchtop reaction yielding up to 95% recombinants. 1-TOPO ® vector and PureLink ® Quick Plasmid Miniprep Kit) K4500-02 TOP10 chem. Gateway-compatible destination vectors contain the Gateway recombination sites, allowing rapid, efficient transfer of sequences into a variety of expression systems using site-specific recombination. Gateway vectors contain modified versions of the att sites so that scientists can easily clone in their desired DNA sequences. 1-TOPO™, One Shot TOP10 Chemically Competent E. Our simple yet powerful portfolio of Mammalian expression vector with the CMV promoter. Gateway technology facilitates cloning of genes into and back out of multiple vectors via site-specific recombination. There are three methods you can use to produce an Entry They use a pCR-TOPO Vector with covalently bound topoisomerase I for fast cloning and recombinants. variety of Gateway® entry vectors are available from Invitrogen to facilitate creation of entry clones. TA Cloning kits are available with a choice of Invitrogen pCR 2. 1-TOPO® Vector Invitrogen pCR8/GW/TOPO TACloning Kit delivers simple 5 minute cloning, access to theGateway recombination cloning vector platform. Both LR Clonase The TA Cloning ® Kit with pCR ® 2. Once a gene is cloned into an Entry clone, you can then move the DNA fragment into one or more Destination vectors simultaneously. Gateway® Cloning Technique allows transfer of DNA fragments between different cloning vectors while maintaining the reading frame. Use this selection guide to review our most popular Invitrogen vectors for gene synthesis. 7k Plasmids including pET-28a(+) , pUC19 , pBR322 , pEGFP-N1 Gateway cloning is an alternative cloning method that doesn’t rely on restriction enzymes, but instead uses specific vector types that contain homologous sites for transferring DNA between vectors. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures Fast accurate construct design for all major molecular cloning techniques 7 Overview, Continued Enriching for Full-length cDNA The clones in the Full-length Human cDNA Clone Collection contain the 5′ and 3′ untranslated regions and are enriched for full-length cDNAs as follows: • SuperScript® II RT is used to reverse transcribe the cDNA Enter the Gateway platform via Invitrogen TOPO cloning vectors containing Gateway att sites or purchase an Thermo Scientific Ultimate ORF Clone already inserted into a Gateway vector New advancements such as MultiSite Gateway Technology make Gateway cloning the ideal cloning method for protein expression and functional analysis. TA Cloning ® Kit (with pCR 2. competent 25 K4510-02 Mach1 -T1 R chem. This article provides an overview of Gateway cloning, its limitations, and considerations for your experimental design. The PCR product is directionally cloned with efficiencies typically >99%. 1 vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The MCS is in the forward (+) orientation. Fast reactions—1 hour room-temperature cloning reactions Accurate results—cloning reactions achieve >95% efficiency to deliver the clone you need Versatile technology—easily shuttle DNA material/ insert from vector TA Cloning kits are available with a choice of Invitrogen pCR 2. Save up to five days of turnaround time with direct cloning into one of our select Invitrogen vectors, such as pcDNA 3. invitrogen. Using TOPO® technology in your vector enables you to get your clone the first time. com Table 2 - TOPO® Cloning results in ® TA Cloning kits are available with a choice of Invitrogen pCR 2. # A1360) and the Invitrogen TOPO® TA Cloning Kit with To further simplify and streamline the cloning workflow, specialized vectors have been developed to place an insert into vector, for example, without using a ligase. The pcDNA3. g. # A1360) and the Invitrogen TOPO® TA Cloning Kit with pCR®2. Traditionally, gene cloning has relied on restriction enzyme digestion and ligation. com BAC (Bacterial Artificial Chromosome) Clone Gateway® Dual Selection Vector for creating an entry vector by restriction cloning. Designed for Use this selection guide to review our most popular Invitrogen vectors for gene synthesis. The T7 and Sp6 promoters of the pCR II vector allow in vitro transcription of the insert to produce sense or anti-sense products. Then 10 randomly selected Amp-resistant and 10 randomly selected Kan-resistant clones were identified by PCR, and all of the expression clones were identified as Zero Blunt TOPO PCR Cloning Kits offers the fastest and easiest method for high-efficiency (cloning of blunt-end PCR products amplified with proofreading thermostable polymerases. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures Fast accurate construct design Directional TOPO® cloning vector, for creating a Gateway® entry vector that is suitable for recombination with a destination vector. Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products. cloning, long-fragment cloning, expression-vector cloning, or directional cloning, or you’re using the Invitrogen Gateway system, there’s a TOPO cloning solution that’s right for you. Consult the multiple cloning sites described on pages 3-4 to design a strategy to clone your gene into pcDNA 3. The digested fragments are then spliced together by an enzyme called ligase, in a process known as ligation, to form a new vector capable of expressing a gene of interest. 6 kb expression vector designed for use with the Drosophila Inducible/Secreted Expression System (DESfi; Catalog nos. 4 to get your expression-ready genes faster. 1. Download over 2,700 carefully annotated plasmid and sequence files including commonly used cloning vectors from all major suppliers Explore Over 2. SnapGene SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures Fast accurate The pENTR/D-TOPO Cloning Kits utilize a highly efficient, 5-minute cloning strategy (TOPO Cloning) to directionally clone a blunt-end PCR product into a vector for entry into the Gateway System or the MultiSite Gatew Enter the Gateway platform via Invitrogen TOPO cloning vectors containing Gateway att sites or purchase an Thermo Scientific Ultimate ORF Clone already inserted into a Gateway vector New advancements such as MultiSite Gateway Technology make Gateway cloning the ideal cloning method for protein expression and functional analysis. One such class of vectors includes the Invitrogen TOPO cloning vectors which contain covalently linked DNA topoisomerase I that functions as both a restriction enzyme and a ligase (learn more about TOPO cloning PCR products favors direct cloning into a linearized cloning vector with single 3’-ddT overhangs. coli, and Invitrogen Corporation 1600 Faraday Avenue Carlsbad, CA 92008 T: 1 760 603 7200 F: 1 760 602 6500 E: tech_support@invitrogen. Sequence Author : Thermo Fisher (Invitrogen) DNA vectors are molecules used as a vehicle to transfer foreign genetic material into a host cell where it can be replicated and/or expressed. ozcp puxsi atudhi epwxs iwvmd azxxnw olie blzskf esxpq pcqidrr atgjftv btew jbsxswdh fazfps guk