t4 ligase rapid kit  (Thermo Fisher)


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    Name:
    Rapid DNA Ligation Kit
    Description:
    Thermo Scientific Rapid DNA Ligation Kit enables fast sticky end or blunt end DNA ligation in only 5 minutes at room temperature The kit contains T4 DNA ligase and a specially formulated 5X rapid ligation buffer optimized for fast and efficient DNA ligation Fast ligation efficiency is equal to that obtained with T4 DNA ligase in a standard 1 hour ligation The ligation reaction mixture can be used directly for bacterial transformation with the TransformAid Bacterial Transformation Kit or with other conventional transformation procedures Highlights• Fast ligation of either sticky or blunt end DNA in only 5 minutes• Convenient reaction mixture can be used directly for bacterial transformationApplications• Routine cloning experiments• Blunt end cloning• Library construction• TA cloningIncludes• T4 DNA Ligase• 5X Rapid Ligation Buffer• Water nuclease free• Detailed ProtocolNotePrior to electroporation it is necessary to inactivate T4 DNA ligase by chloroform extraction or spin column purification e g with Thermo Scientific GeneJET PCR Purification Kit K0701 Related ProductsRapid DNA Ligation Kit
    Catalog Number:
    k1422
    Price:
    None
    Applications:
    Cloning|PCR Cloning|Restriction Enzyme Cloning
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher t4 ligase rapid kit
    Thermo Scientific Rapid DNA Ligation Kit enables fast sticky end or blunt end DNA ligation in only 5 minutes at room temperature The kit contains T4 DNA ligase and a specially formulated 5X rapid ligation buffer optimized for fast and efficient DNA ligation Fast ligation efficiency is equal to that obtained with T4 DNA ligase in a standard 1 hour ligation The ligation reaction mixture can be used directly for bacterial transformation with the TransformAid Bacterial Transformation Kit or with other conventional transformation procedures Highlights• Fast ligation of either sticky or blunt end DNA in only 5 minutes• Convenient reaction mixture can be used directly for bacterial transformationApplications• Routine cloning experiments• Blunt end cloning• Library construction• TA cloningIncludes• T4 DNA Ligase• 5X Rapid Ligation Buffer• Water nuclease free• Detailed ProtocolNotePrior to electroporation it is necessary to inactivate T4 DNA ligase by chloroform extraction or spin column purification e g with Thermo Scientific GeneJET PCR Purification Kit K0701 Related ProductsRapid DNA Ligation Kit
    https://www.bioz.com/result/t4 ligase rapid kit/product/Thermo Fisher
    Average 96 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    t4 ligase rapid kit - by Bioz Stars, 2020-05
    96/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Syntaxin-17 delivers PINK1/parkin-dependent mitochondrial vesicles to the endolysosomal system
    Article Snippet: .. Purified components were ligated using Rapid DNA Ligation kit (Thermo Fisher Scientific) according to the manufacturer’s instructions, and positive clones were identified by sequencing. .. GFP-Stx17 was constructed by double-digesting YFP-Stx17 and pEGFP-C1 (Takara Bio Inc.) with XhoI and XbaI, followed by a ligation as above.

    Amplification:

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Bioluminescent Imaging of Genetically Selected Induced Pluripotent Stem Cell-Derived Cardiomyocytes after Transplantation into Infarcted Heart of Syngeneic Recipients
    Article Snippet: .. UbC promoter (pUbC) was amplified from plasmid pLenti6/UbC/V5-DEST (Invitrogen, Life Technologies, Carlsbad, CA) with Dream Taq PCR Master Mix (Fermentas, Thermo Scientific, Waltham, MA) using primers flanked by restriction sites for Xho I and Hind III (Fermentas). pGL4.14 [luc2 /Hygro] plasmid and the PCR-product were digested with both enzymes, purified with NucleoSpin Gel and PCR Clean-up-Kit (Macherey-Nagel, Düren, Germany) and ligated overnight at 4°C with Rapid DNA Ligation Kit (Fermentas) to yield the pGL4.14-pUbC [luc2 /Hygro] vector. .. The PGK promoter was removed from the pPPGKIP vector by digestion with Sal I and Hind III and subcloned between the XhoI and HindIII cutting sites of the pGL4.14 [luc2 /Hygro] plasmid to obtain pGL4.14-pPGK [luc2 /Hygro] vector.

    DNA Ligation:

    Article Title: The Novel Candida albicans Transporter Dur31 Is a Multi-Stage Pathogenicity Factor
    Article Snippet: .. The DUR31 insert and CIp10 vector were then ligated for 30 min at 22°C using the Rapid DNA Ligation Kit (Fermentas). .. Five µl of ligation product was used for the transformation of E. coli DH5α and positive clones were selected on LB agar plates supplemented with 50 µg ml−1 Ampicillin.

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Syntaxin-17 delivers PINK1/parkin-dependent mitochondrial vesicles to the endolysosomal system
    Article Snippet: .. Purified components were ligated using Rapid DNA Ligation kit (Thermo Fisher Scientific) according to the manufacturer’s instructions, and positive clones were identified by sequencing. .. GFP-Stx17 was constructed by double-digesting YFP-Stx17 and pEGFP-C1 (Takara Bio Inc.) with XhoI and XbaI, followed by a ligation as above.

    Article Title: Production of four Neurospora crassa lytic polysaccharide monooxygenases in Pichia pastoris monitored by a fluorimetric assay
    Article Snippet: .. Construction of pmo expression vectors for P. pastoris The synthetic pmo genes were digested with the restriction enzymes Bst BI and Xba I and ligated into the equally treated vector pPICZα A using the Rapid DNA Ligation Kit from Fermentas. .. The procedures resulted in plasmids carrying genes encoding proteins with their native signal sequences cloned under the control of the methanol inducible AOX1 promoter.

    Article Title: A combination of transposable elements and magnetic cell sorting provides a very efficient transgenesis system for chicken primary erythroid progenitors
    Article Snippet: .. Considering that a 4 bp recognition site will occur roughly every 256 bp, and therefore that 200 ng of Tai I-digested gDNA correspond to 1.28 pmol DNA, 200 ng of digested DNA were ligated to a 3× molar excess of the annealed splinkerette adaptor (Splinklong-ACGT+Splinkshort-hairpin) with the T4 DNA ligase (4 U), for 1 hour at 22°C, using the Fermentas Rapid DNA Ligation kit. .. In order to improve the ligation yield, digested DNA was heated for 5 min at 60°C before being ligated.

    Article Title: Bioluminescent Imaging of Genetically Selected Induced Pluripotent Stem Cell-Derived Cardiomyocytes after Transplantation into Infarcted Heart of Syngeneic Recipients
    Article Snippet: .. UbC promoter (pUbC) was amplified from plasmid pLenti6/UbC/V5-DEST (Invitrogen, Life Technologies, Carlsbad, CA) with Dream Taq PCR Master Mix (Fermentas, Thermo Scientific, Waltham, MA) using primers flanked by restriction sites for Xho I and Hind III (Fermentas). pGL4.14 [luc2 /Hygro] plasmid and the PCR-product were digested with both enzymes, purified with NucleoSpin Gel and PCR Clean-up-Kit (Macherey-Nagel, Düren, Germany) and ligated overnight at 4°C with Rapid DNA Ligation Kit (Fermentas) to yield the pGL4.14-pUbC [luc2 /Hygro] vector. .. The PGK promoter was removed from the pPPGKIP vector by digestion with Sal I and Hind III and subcloned between the XhoI and HindIII cutting sites of the pGL4.14 [luc2 /Hygro] plasmid to obtain pGL4.14-pPGK [luc2 /Hygro] vector.

    Article Title: FlgM as a Secretion Moiety for the Development of an Inducible Type III Secretion System
    Article Snippet: .. Rapid DNA ligation kit, Pfu DNA polymerase and 10× MgSO4 -PCR buffer were obtained from Fermentas. .. Tris-Glycine gels were purchased from Invitrogen and Biorad.

    Purification:

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Syntaxin-17 delivers PINK1/parkin-dependent mitochondrial vesicles to the endolysosomal system
    Article Snippet: .. Purified components were ligated using Rapid DNA Ligation kit (Thermo Fisher Scientific) according to the manufacturer’s instructions, and positive clones were identified by sequencing. .. GFP-Stx17 was constructed by double-digesting YFP-Stx17 and pEGFP-C1 (Takara Bio Inc.) with XhoI and XbaI, followed by a ligation as above.

    Article Title: Bioluminescent Imaging of Genetically Selected Induced Pluripotent Stem Cell-Derived Cardiomyocytes after Transplantation into Infarcted Heart of Syngeneic Recipients
    Article Snippet: .. UbC promoter (pUbC) was amplified from plasmid pLenti6/UbC/V5-DEST (Invitrogen, Life Technologies, Carlsbad, CA) with Dream Taq PCR Master Mix (Fermentas, Thermo Scientific, Waltham, MA) using primers flanked by restriction sites for Xho I and Hind III (Fermentas). pGL4.14 [luc2 /Hygro] plasmid and the PCR-product were digested with both enzymes, purified with NucleoSpin Gel and PCR Clean-up-Kit (Macherey-Nagel, Düren, Germany) and ligated overnight at 4°C with Rapid DNA Ligation Kit (Fermentas) to yield the pGL4.14-pUbC [luc2 /Hygro] vector. .. The PGK promoter was removed from the pPPGKIP vector by digestion with Sal I and Hind III and subcloned between the XhoI and HindIII cutting sites of the pGL4.14 [luc2 /Hygro] plasmid to obtain pGL4.14-pPGK [luc2 /Hygro] vector.

    Polymerase Chain Reaction:

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Bioluminescent Imaging of Genetically Selected Induced Pluripotent Stem Cell-Derived Cardiomyocytes after Transplantation into Infarcted Heart of Syngeneic Recipients
    Article Snippet: .. UbC promoter (pUbC) was amplified from plasmid pLenti6/UbC/V5-DEST (Invitrogen, Life Technologies, Carlsbad, CA) with Dream Taq PCR Master Mix (Fermentas, Thermo Scientific, Waltham, MA) using primers flanked by restriction sites for Xho I and Hind III (Fermentas). pGL4.14 [luc2 /Hygro] plasmid and the PCR-product were digested with both enzymes, purified with NucleoSpin Gel and PCR Clean-up-Kit (Macherey-Nagel, Düren, Germany) and ligated overnight at 4°C with Rapid DNA Ligation Kit (Fermentas) to yield the pGL4.14-pUbC [luc2 /Hygro] vector. .. The PGK promoter was removed from the pPPGKIP vector by digestion with Sal I and Hind III and subcloned between the XhoI and HindIII cutting sites of the pGL4.14 [luc2 /Hygro] plasmid to obtain pGL4.14-pPGK [luc2 /Hygro] vector.

    Article Title: FlgM as a Secretion Moiety for the Development of an Inducible Type III Secretion System
    Article Snippet: .. Rapid DNA ligation kit, Pfu DNA polymerase and 10× MgSO4 -PCR buffer were obtained from Fermentas. .. Tris-Glycine gels were purchased from Invitrogen and Biorad.

    Expressing:

    Article Title: Production of four Neurospora crassa lytic polysaccharide monooxygenases in Pichia pastoris monitored by a fluorimetric assay
    Article Snippet: .. Construction of pmo expression vectors for P. pastoris The synthetic pmo genes were digested with the restriction enzymes Bst BI and Xba I and ligated into the equally treated vector pPICZα A using the Rapid DNA Ligation Kit from Fermentas. .. The procedures resulted in plasmids carrying genes encoding proteins with their native signal sequences cloned under the control of the methanol inducible AOX1 promoter.

    Sequencing:

    Article Title: Syntaxin-17 delivers PINK1/parkin-dependent mitochondrial vesicles to the endolysosomal system
    Article Snippet: .. Purified components were ligated using Rapid DNA Ligation kit (Thermo Fisher Scientific) according to the manufacturer’s instructions, and positive clones were identified by sequencing. .. GFP-Stx17 was constructed by double-digesting YFP-Stx17 and pEGFP-C1 (Takara Bio Inc.) with XhoI and XbaI, followed by a ligation as above.

    Plasmid Preparation:

    Article Title: The Novel Candida albicans Transporter Dur31 Is a Multi-Stage Pathogenicity Factor
    Article Snippet: .. The DUR31 insert and CIp10 vector were then ligated for 30 min at 22°C using the Rapid DNA Ligation Kit (Fermentas). .. Five µl of ligation product was used for the transformation of E. coli DH5α and positive clones were selected on LB agar plates supplemented with 50 µg ml−1 Ampicillin.

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Production of four Neurospora crassa lytic polysaccharide monooxygenases in Pichia pastoris monitored by a fluorimetric assay
    Article Snippet: .. Construction of pmo expression vectors for P. pastoris The synthetic pmo genes were digested with the restriction enzymes Bst BI and Xba I and ligated into the equally treated vector pPICZα A using the Rapid DNA Ligation Kit from Fermentas. .. The procedures resulted in plasmids carrying genes encoding proteins with their native signal sequences cloned under the control of the methanol inducible AOX1 promoter.

    Article Title: Transfection of Peripheral Blood Monocytes with SOX2 Enhances Multipotency, Proliferation, and Redifferentiation into Neohepatocytes and Insulin-Producing Cells
    Article Snippet: .. Both the insert and vector were ligated using T4-Ligase Rapid kit (Fermentas). .. Ligation product was transformed overnight in DH5a E. coli , and the SOX2 clones were then isolated using Fermentas jet plasmid miniprep.

    Article Title: Bioluminescent Imaging of Genetically Selected Induced Pluripotent Stem Cell-Derived Cardiomyocytes after Transplantation into Infarcted Heart of Syngeneic Recipients
    Article Snippet: .. UbC promoter (pUbC) was amplified from plasmid pLenti6/UbC/V5-DEST (Invitrogen, Life Technologies, Carlsbad, CA) with Dream Taq PCR Master Mix (Fermentas, Thermo Scientific, Waltham, MA) using primers flanked by restriction sites for Xho I and Hind III (Fermentas). pGL4.14 [luc2 /Hygro] plasmid and the PCR-product were digested with both enzymes, purified with NucleoSpin Gel and PCR Clean-up-Kit (Macherey-Nagel, Düren, Germany) and ligated overnight at 4°C with Rapid DNA Ligation Kit (Fermentas) to yield the pGL4.14-pUbC [luc2 /Hygro] vector. .. The PGK promoter was removed from the pPPGKIP vector by digestion with Sal I and Hind III and subcloned between the XhoI and HindIII cutting sites of the pGL4.14 [luc2 /Hygro] plasmid to obtain pGL4.14-pPGK [luc2 /Hygro] vector.

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  • 96
    Thermo Fisher rapid dna ligation kit
    Rapid Dna Ligation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rapid dna ligation kit/product/Thermo Fisher
    Average 96 stars, based on 27 article reviews
    Price from $9.99 to $1999.99
    rapid dna ligation kit - by Bioz Stars, 2020-05
    96/100 stars
      Buy from Supplier

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