pentr directional topo cloning kit  (Thermo Fisher)


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    Name:
    pENTR TEV D TOPO Cloning Kit
    Description:
    The pENTR TEV 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 Gateway System Blunt end PCR products clone directionally into the pENTR TEV D TOPO entry vector at greater than 90 efficiency and include a TEV protease dependent cleavage site for removal of N terminal tags from expressed proteins The kit comes with everything necessary to clone and select your PCR amplified gene of interest • Gateway System Ready Rapidly shuttle cloned genes between multiple vector systems• Fast and Easy Go from PCR to Gateway Entry clone in just 3 steps and as little as 5 minutes hands on time• Efficient Achieve over 90 clones with correct insert in the right direction• Proven Reliable performance for over a decade pENTR TEV D TOPO Cloning Kits Overview VECTOR pENTR TEV D TOPO Vector Directional cloning vector for entry to the Gateway System optimized with a TEV cleavage site for removal of N terminal tags CLONING METHOD Directional TOPO Cloning Topoisomerase I based 5 minute directional ligation of blunt end proofreading polymerase amplified PCR products to the vector COMPETENT CELLS Two Options Choose from kits with either high efficiency or fast growing competent cells Simple Access to the Gateway SystemFor access to the Gateway System just PCR amplify your gene of interest and add the product straight to the provided topoisomerase charged pENTR SD D TOPO vector incubate 5 minutes and transform the provided competent E coli cells The resulting attL containing Gateway Entry clones are ready for efficient recombination with your choice of Gateway Destination vectors Optimized pENTR TEV D TOPO VectorThe pENTR TEV D TOPO vector includes a Tobacco Etch Virus TEV recognition site for protease dependent cleavage of an N terminal tag from your recombinant protein The vector has M13 and T7 primer sequencing sites and attL recombination sites flanking the PCR product insertion site This allows clones to be easily sequence verified and recombined into your choice of attR containing Gateway destination vectors A Kanamycin resistance gene and a pUC origin are used for selection and high copy propagation in E coli Simplified Directional CloningWith Directional TOPO cloning technology there is no need for PCR clean up vector preparation or other time intensive DNA manipulation steps Just add your PCR reaction straight to the provided topoisomerase charged vector incubate 5 minutes transform and obtain up to 90 directionally inserted clones A four base overhang on the vector pairs with a four base sequence designed into the forward primer used in your PCR reaction to provide directionality to the topoisomerase ligation reaction Figure 2 The Power of Gateway Recombination Cloning TechnologyGateway recombination cloning technology circumvents the limitations of restriction mediated cloning enabling you to access virtually any expression system in a simple one hour 99 efficient and reversible Gateway recombination reaction The ability to move the same sequence of DNA between different vectors without using restriction enzymes ligase subcloning steps screening of countless colonies or re sequencing will help save you time money and effort Leading Cloning TechnologiesWhen it comes to cloning TOPO cloning technology and Gateway recombination cloning technology have been a reliable partner for thousands of scientists for over ten years Fast simple to use and efficient TOPO cloning and Gateway recombination allow for rapid cloning and subsequent transferring of genes between a wide assortment of Gateway expression vectors Kit OptionsThe pENTR TEV D TOPO Cloning Kit can be purchased with either TOP10 competent cells for standard cloning or Mach1 T1R competent cells for fast growth For Research Use Only Not intended for use in animal or human therapeutic or diagnostic use
    Catalog Number:
    k252520
    Price:
    None
    Applications:
    Cloning|Entry Vectors & Kits|Gateway Cloning|pENTR Vectors & Kits
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
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    Structured Review

    Thermo Fisher pentr directional topo cloning kit
    The pENTR TEV 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 Gateway System Blunt end PCR products clone directionally into the pENTR TEV D TOPO entry vector at greater than 90 efficiency and include a TEV protease dependent cleavage site for removal of N terminal tags from expressed proteins The kit comes with everything necessary to clone and select your PCR amplified gene of interest • Gateway System Ready Rapidly shuttle cloned genes between multiple vector systems• Fast and Easy Go from PCR to Gateway Entry clone in just 3 steps and as little as 5 minutes hands on time• Efficient Achieve over 90 clones with correct insert in the right direction• Proven Reliable performance for over a decade pENTR TEV D TOPO Cloning Kits Overview VECTOR pENTR TEV D TOPO Vector Directional cloning vector for entry to the Gateway System optimized with a TEV cleavage site for removal of N terminal tags CLONING METHOD Directional TOPO Cloning Topoisomerase I based 5 minute directional ligation of blunt end proofreading polymerase amplified PCR products to the vector COMPETENT CELLS Two Options Choose from kits with either high efficiency or fast growing competent cells Simple Access to the Gateway SystemFor access to the Gateway System just PCR amplify your gene of interest and add the product straight to the provided topoisomerase charged pENTR SD D TOPO vector incubate 5 minutes and transform the provided competent E coli cells The resulting attL containing Gateway Entry clones are ready for efficient recombination with your choice of Gateway Destination vectors Optimized pENTR TEV D TOPO VectorThe pENTR TEV D TOPO vector includes a Tobacco Etch Virus TEV recognition site for protease dependent cleavage of an N terminal tag from your recombinant protein The vector has M13 and T7 primer sequencing sites and attL recombination sites flanking the PCR product insertion site This allows clones to be easily sequence verified and recombined into your choice of attR containing Gateway destination vectors A Kanamycin resistance gene and a pUC origin are used for selection and high copy propagation in E coli Simplified Directional CloningWith Directional TOPO cloning technology there is no need for PCR clean up vector preparation or other time intensive DNA manipulation steps Just add your PCR reaction straight to the provided topoisomerase charged vector incubate 5 minutes transform and obtain up to 90 directionally inserted clones A four base overhang on the vector pairs with a four base sequence designed into the forward primer used in your PCR reaction to provide directionality to the topoisomerase ligation reaction Figure 2 The Power of Gateway Recombination Cloning TechnologyGateway recombination cloning technology circumvents the limitations of restriction mediated cloning enabling you to access virtually any expression system in a simple one hour 99 efficient and reversible Gateway recombination reaction The ability to move the same sequence of DNA between different vectors without using restriction enzymes ligase subcloning steps screening of countless colonies or re sequencing will help save you time money and effort Leading Cloning TechnologiesWhen it comes to cloning TOPO cloning technology and Gateway recombination cloning technology have been a reliable partner for thousands of scientists for over ten years Fast simple to use and efficient TOPO cloning and Gateway recombination allow for rapid cloning and subsequent transferring of genes between a wide assortment of Gateway expression vectors Kit OptionsThe pENTR TEV D TOPO Cloning Kit can be purchased with either TOP10 competent cells for standard cloning or Mach1 T1R competent cells for fast growth For Research Use Only Not intended for use in animal or human therapeutic or diagnostic use
    https://www.bioz.com/result/pentr directional topo cloning kit/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    pentr directional topo cloning kit - by Bioz Stars, 2021-01
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Masking of a Nuclear Signal Motif by Monoubiquitination Leads to Mislocalization and Degradation of the Regulatory Enzyme Cytidylyltransferase ▿Masking of a Nuclear Signal Motif by Monoubiquitination Leads to Mislocalization and Degradation of the Regulatory Enzyme Cytidylyltransferase ▿ †
    Article Snippet: .. LysoTracker Red, mouse monoclonal V5 antibody, the To-Pro-3 nuclear staining kit, the pcDNA3.1D cloning kit, Escherichia coli OneShot competent cells, the pENTR directional TOPO cloning kits, and the Gateway mammalian expression system were purchased from Invitrogen (Carlsbad, CA). .. BD Talon purification and buffer kits were purchased from BD Biosciences (San Jose, CA).

    Article Title: Anaplasma phagocytophilum Outer Membrane Protein A Interacts with Sialylated Glycoproteins To Promote Infection of Mammalian Host Cells
    Article Snippet: .. Amplicons were cloned into pENTR/TEV/D-TOPO (Invitrogen) as described previously ( ) to yield pENTR-candidate gene entry plasmids containing the genes of interest. .. Plasmid inserts were verified, and recombination of the candidate gene insert downstream of and in frame with the gene encoding GST was achieved using the pDest-15 vector (Invitrogen) as described previously ( ).

    Article Title: Functional Conservation in the SIAMESE-RELATED Family of Cyclin-Dependent Kinase Inhibitors in Land Plants
    Article Snippet: .. Coding regions of SMRs were synthesized by Integrated DNA Technologies, PCR amplified with appropriate primers , and inserted into the vector (pENTR/D-TOPO) using a pENTR Directional TOPO Cloning Kit (Life Technologies). .. Error-free entry clones were confirmed by sequence analysis.

    Article Title: A miR-34a-SIRT6 axis in the squamous cell differentiation network
    Article Snippet: .. After sequence verification, the coding sequence of p53R248W or SIRT6 was transferred by recombination into the destination lentiviral vector pINDUCER-20 ( ) using the pENTR™ Directional TOPO® Cloning kit (Invitrogen). .. Cells stably infected with these lentiviruses (after G418 selection (500 μg/ml)) were induced to express p53R248W or SIRT6 by the presence of doxycycline in the culture medium.

    Article Title: Sequencing genomes from single cells by polymerase cloning.
    Article Snippet: .. Genome sequencing currently requires DNA from pools of numerous nearly identical cells (clones), leaving the genome sequences of many difficult-to-culture microorganisms unattainable. .. Genome sequencing currently requires DNA from pools of numerous nearly identical cells (clones), leaving the genome sequences of many difficult-to-culture microorganisms unattainable.

    Article Title: Flexibility and constraint: Evolutionary remodeling of the sporulation initiation pathway in Firmicutes
    Article Snippet: .. Briefly, the nucleotide sequences described above were cloned into pENTR/D-TOPO entry vectors using the pENTR Directional TOPO Cloning Kit (Thermo Fisher Scientific). .. The coding sequences were subsequently transferred to destination vectors using the Gateway LR reaction (Thermo Fisher Scientific), yielding expression plasmids encoding affinity-tagged proteins under control of an IPTG-inducible promoter.

    Article Title: LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25 [OPEN]
    Article Snippet: .. To construct the Pro35S : LUH-myc ( LUH-myc ) plasmid, the coding sequence of LUH was PCR amplified, cloned into pENTR using a pENTR Directional TOPO Cloning Kit (Invitrogen), and recombined with the binary vector PGWB17 . .. These constructs were transformed into Agrobacterium tumefaciens strain GV3101 and then the transformed GV3101 cells were used to generate transgenic Arabidopsis plants using the floral dip method ( ).

    Article Title: Overexpression of OsF3H modulates WBPH stress by alteration of phenylpropanoid pathway at a transcriptomic and metabolomic level in Oryza sativa
    Article Snippet: .. Entry clones were generated by inserting template DNA into pENTR/D-TOPO cloning vectors, following the manufacturer’s instructions [for details, see the user manual for the pENTR Directional TOPO cloning kit (Invitrogen)]. .. The TOPO cloning reaction was transformed into DH5α cells using heat shock, and spread on LB media containing kanamycin as a selection marker.

    Amplification:

    Article Title: Functional Conservation in the SIAMESE-RELATED Family of Cyclin-Dependent Kinase Inhibitors in Land Plants
    Article Snippet: .. Coding regions of SMRs were synthesized by Integrated DNA Technologies, PCR amplified with appropriate primers , and inserted into the vector (pENTR/D-TOPO) using a pENTR Directional TOPO Cloning Kit (Life Technologies). .. Error-free entry clones were confirmed by sequence analysis.

    Article Title: LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25 [OPEN]
    Article Snippet: .. To construct the Pro35S : LUH-myc ( LUH-myc ) plasmid, the coding sequence of LUH was PCR amplified, cloned into pENTR using a pENTR Directional TOPO Cloning Kit (Invitrogen), and recombined with the binary vector PGWB17 . .. These constructs were transformed into Agrobacterium tumefaciens strain GV3101 and then the transformed GV3101 cells were used to generate transgenic Arabidopsis plants using the floral dip method ( ).

    Synthesized:

    Article Title: Functional Conservation in the SIAMESE-RELATED Family of Cyclin-Dependent Kinase Inhibitors in Land Plants
    Article Snippet: .. Coding regions of SMRs were synthesized by Integrated DNA Technologies, PCR amplified with appropriate primers , and inserted into the vector (pENTR/D-TOPO) using a pENTR Directional TOPO Cloning Kit (Life Technologies). .. Error-free entry clones were confirmed by sequence analysis.

    Construct:

    Article Title: LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25 [OPEN]
    Article Snippet: .. To construct the Pro35S : LUH-myc ( LUH-myc ) plasmid, the coding sequence of LUH was PCR amplified, cloned into pENTR using a pENTR Directional TOPO Cloning Kit (Invitrogen), and recombined with the binary vector PGWB17 . .. These constructs were transformed into Agrobacterium tumefaciens strain GV3101 and then the transformed GV3101 cells were used to generate transgenic Arabidopsis plants using the floral dip method ( ).

    Polymerase Chain Reaction:

    Article Title: Functional Conservation in the SIAMESE-RELATED Family of Cyclin-Dependent Kinase Inhibitors in Land Plants
    Article Snippet: .. Coding regions of SMRs were synthesized by Integrated DNA Technologies, PCR amplified with appropriate primers , and inserted into the vector (pENTR/D-TOPO) using a pENTR Directional TOPO Cloning Kit (Life Technologies). .. Error-free entry clones were confirmed by sequence analysis.

    Article Title: LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25 [OPEN]
    Article Snippet: .. To construct the Pro35S : LUH-myc ( LUH-myc ) plasmid, the coding sequence of LUH was PCR amplified, cloned into pENTR using a pENTR Directional TOPO Cloning Kit (Invitrogen), and recombined with the binary vector PGWB17 . .. These constructs were transformed into Agrobacterium tumefaciens strain GV3101 and then the transformed GV3101 cells were used to generate transgenic Arabidopsis plants using the floral dip method ( ).

    Generated:

    Article Title: Overexpression of OsF3H modulates WBPH stress by alteration of phenylpropanoid pathway at a transcriptomic and metabolomic level in Oryza sativa
    Article Snippet: .. Entry clones were generated by inserting template DNA into pENTR/D-TOPO cloning vectors, following the manufacturer’s instructions [for details, see the user manual for the pENTR Directional TOPO cloning kit (Invitrogen)]. .. The TOPO cloning reaction was transformed into DH5α cells using heat shock, and spread on LB media containing kanamycin as a selection marker.

    Expressing:

    Article Title: Masking of a Nuclear Signal Motif by Monoubiquitination Leads to Mislocalization and Degradation of the Regulatory Enzyme Cytidylyltransferase ▿Masking of a Nuclear Signal Motif by Monoubiquitination Leads to Mislocalization and Degradation of the Regulatory Enzyme Cytidylyltransferase ▿ †
    Article Snippet: .. LysoTracker Red, mouse monoclonal V5 antibody, the To-Pro-3 nuclear staining kit, the pcDNA3.1D cloning kit, Escherichia coli OneShot competent cells, the pENTR directional TOPO cloning kits, and the Gateway mammalian expression system were purchased from Invitrogen (Carlsbad, CA). .. BD Talon purification and buffer kits were purchased from BD Biosciences (San Jose, CA).

    Sequencing:

    Article Title: A miR-34a-SIRT6 axis in the squamous cell differentiation network
    Article Snippet: .. After sequence verification, the coding sequence of p53R248W or SIRT6 was transferred by recombination into the destination lentiviral vector pINDUCER-20 ( ) using the pENTR™ Directional TOPO® Cloning kit (Invitrogen). .. Cells stably infected with these lentiviruses (after G418 selection (500 μg/ml)) were induced to express p53R248W or SIRT6 by the presence of doxycycline in the culture medium.

    Article Title: LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25 [OPEN]
    Article Snippet: .. To construct the Pro35S : LUH-myc ( LUH-myc ) plasmid, the coding sequence of LUH was PCR amplified, cloned into pENTR using a pENTR Directional TOPO Cloning Kit (Invitrogen), and recombined with the binary vector PGWB17 . .. These constructs were transformed into Agrobacterium tumefaciens strain GV3101 and then the transformed GV3101 cells were used to generate transgenic Arabidopsis plants using the floral dip method ( ).

    Staining:

    Article Title: Masking of a Nuclear Signal Motif by Monoubiquitination Leads to Mislocalization and Degradation of the Regulatory Enzyme Cytidylyltransferase ▿Masking of a Nuclear Signal Motif by Monoubiquitination Leads to Mislocalization and Degradation of the Regulatory Enzyme Cytidylyltransferase ▿ †
    Article Snippet: .. LysoTracker Red, mouse monoclonal V5 antibody, the To-Pro-3 nuclear staining kit, the pcDNA3.1D cloning kit, Escherichia coli OneShot competent cells, the pENTR directional TOPO cloning kits, and the Gateway mammalian expression system were purchased from Invitrogen (Carlsbad, CA). .. BD Talon purification and buffer kits were purchased from BD Biosciences (San Jose, CA).

    Plasmid Preparation:

    Article Title: Functional Conservation in the SIAMESE-RELATED Family of Cyclin-Dependent Kinase Inhibitors in Land Plants
    Article Snippet: .. Coding regions of SMRs were synthesized by Integrated DNA Technologies, PCR amplified with appropriate primers , and inserted into the vector (pENTR/D-TOPO) using a pENTR Directional TOPO Cloning Kit (Life Technologies). .. Error-free entry clones were confirmed by sequence analysis.

    Article Title: A miR-34a-SIRT6 axis in the squamous cell differentiation network
    Article Snippet: .. After sequence verification, the coding sequence of p53R248W or SIRT6 was transferred by recombination into the destination lentiviral vector pINDUCER-20 ( ) using the pENTR™ Directional TOPO® Cloning kit (Invitrogen). .. Cells stably infected with these lentiviruses (after G418 selection (500 μg/ml)) were induced to express p53R248W or SIRT6 by the presence of doxycycline in the culture medium.

    Article Title: LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25 [OPEN]
    Article Snippet: .. To construct the Pro35S : LUH-myc ( LUH-myc ) plasmid, the coding sequence of LUH was PCR amplified, cloned into pENTR using a pENTR Directional TOPO Cloning Kit (Invitrogen), and recombined with the binary vector PGWB17 . .. These constructs were transformed into Agrobacterium tumefaciens strain GV3101 and then the transformed GV3101 cells were used to generate transgenic Arabidopsis plants using the floral dip method ( ).

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  • 99
    Thermo Fisher pentr directional topo cloning kit
    Pentr Directional Topo Cloning Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pentr directional topo cloning kit/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    pentr directional topo cloning kit - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

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