t4 dna ligase 5 u µl  (Thermo Fisher)


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    Structured Review

    Thermo Fisher t4 dna ligase 5 u µl
    T4 Dna Ligase 5 U µl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase 5 u µl/product/Thermo Fisher
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t4 dna ligase 5 u µl - by Bioz Stars, 2020-04
    95/100 stars

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    Related Articles

    Methylation Sequencing:

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: Paragraph title: Bisulfite sequencing PCR ... The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA).

    Clone Assay:

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: .. The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA). .. Ten positive clones were picked for DNA sequencing analysis.

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: This was done according to the cloning strategy described in Ran et al ( ). .. The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014).

    Amplification:

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: .. The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA). .. Ten positive clones were picked for DNA sequencing analysis.

    Agarose Gel Electrophoresis:

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature. .. Agarose gel electrophoresis: 5 μl of the solution containing the DNA fragment(s) to be analysed was mixed with 1 μl of gel loading dye (cat. # R0611; Thermo Fisher Scientific) and analysed on a 1% (w/v) agarose gel (cat. # BN‐50004; BioNordika Denmark A/S, Herlev, Denmark) in TAE buffer [1 mM EDTA (cat. # E6758; Sigma‐Aldrich Corp.), 40 mM Tris (cat. # T1503; Sigma‐Aldrich Corp.) and 20 mM acetic acid (cat. # 33209; Sigma‐Aldrich Corp.)].

    Modification:

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: Genomic DNA was extracted from mouse BMDM and modified using the the EpiTect Fast DNA Bisulfite Kit (59824, QIAGEN, Dusseldorf, German). .. The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA).

    Methylation:

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA). .. The level of methylated CpG sites between −96 bp to +188 bp on the ABCG1 promotor were compared using genomic DNA from ADK KO and ADK WT BMDMs.

    Mutagenesis:

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: Site‐directed mutagenesis: 45 μl of the PCR product is treated with DpnI (cat. # FD1703; Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's instructions to digest the template DNA used for the amplifications. .. An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature.

    Ligation:

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: .. An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature. ..

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014). .. Afterwards, the digested ligation reaction was transformed into chemically competent E. coli bacteria.

    Plasmid Preparation:

    Article Title: Multiplex mutagenesis of four clustered CrRLK1L with CRISPR/Cas9 exposes their growth regulatory roles in response to metal ions
    Article Snippet: .. For the Golden Gate reaction, 150 ng of the destination vector (pGGZ003) and 250 ng of each entry vector containing the promoter, Cas9, terminator, gRNA1, and BASTA resistance were mixed with 250 ng each of the gRNA2 and gRNA3 PCR products, 0.2 µL BSA protein (New England Biolabs, B9000S), 2 µL ligase buffer, 1.2 µL T4 DNA ligase (ThermoFisher, #EL0011), 1 µL BsaI (NEB, #R0535S) and distilled water (to 20 µL). .. To eliminate not fully-ligated intermediate products, we added 0.85 µL ATP (25 mM) and 1 µL Plasmid-Safe ATP-dependent DNase (Epicentre, E3101K) and incubated for 60 min at 37 °C and for 30 min at 70 °C.

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: .. The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA). .. Ten positive clones were picked for DNA sequencing analysis.

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: General procedures Plasmid purification: The Nucleospin Plasmid EasyPure Kit (cat. # 740727.250; Macherey‐Nagel GmbH & Co. KG, Düren, Germany) was used for plasmid purification according to the manufacturer's instructions. .. An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature.

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: .. The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014). .. To digest any residual linearized DNA, a digestion with Plasmid‐Safe exonuclease (10 U/μl; Biozym) was performed.

    Construct:

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: For the generation of each enhancer deletion site, a flanking pair of sgRNA constructs was synthesized as DNA oligonucleotides (Eurofins, HPSF, no modifications). .. The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014).

    Purification:

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: General procedures Plasmid purification: The Nucleospin Plasmid EasyPure Kit (cat. # 740727.250; Macherey‐Nagel GmbH & Co. KG, Düren, Germany) was used for plasmid purification according to the manufacturer's instructions. .. An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature.

    HAC Assay:

    Article Title: Genome-wide mapping of nucleotide excision repair with XR-seq
    Article Snippet: Glycogen (Roche, cat. no. 10901393001) Sodium acetate (NaAc; Amresco, cat. no. 0602–500G) Acetic acid (HAc; VWR, cat. no. BDH3094) CAUTION HAc is an irritant and is flammable. .. T4 DNA ligase (5 U/μl; Thermo Fisher, cat no. 15224041) PEG 8000 (50% (wt/vol), from T4 RNA ligase reaction buffer; NEB, cat. no. B0216L) Anti-(6–4)PP (Cosmo Bio, cat. no. CAC-NM-DND-002) CRITICAL Antibodies from other suppliers may not be as efficient.

    Concentration Assay:

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: The top and bottom strands of sgRNA encoding oligonucleotide (final concentration of 100 μM) were annealed and phosphorylated by polynucleotide kinase (T4 PNK, Thermo Fisher scientific, #EK0031) reaction. .. The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014).

    Incubation:

    Article Title: Multiplex mutagenesis of four clustered CrRLK1L with CRISPR/Cas9 exposes their growth regulatory roles in response to metal ions
    Article Snippet: For the Golden Gate reaction, 150 ng of the destination vector (pGGZ003) and 250 ng of each entry vector containing the promoter, Cas9, terminator, gRNA1, and BASTA resistance were mixed with 250 ng each of the gRNA2 and gRNA3 PCR products, 0.2 µL BSA protein (New England Biolabs, B9000S), 2 µL ligase buffer, 1.2 µL T4 DNA ligase (ThermoFisher, #EL0011), 1 µL BsaI (NEB, #R0535S) and distilled water (to 20 µL). .. To eliminate not fully-ligated intermediate products, we added 0.85 µL ATP (25 mM) and 1 µL Plasmid-Safe ATP-dependent DNase (Epicentre, E3101K) and incubated for 60 min at 37 °C and for 30 min at 70 °C.

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: .. An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature. ..

    Polymerase Chain Reaction:

    Article Title: Multiplex mutagenesis of four clustered CrRLK1L with CRISPR/Cas9 exposes their growth regulatory roles in response to metal ions
    Article Snippet: .. For the Golden Gate reaction, 150 ng of the destination vector (pGGZ003) and 250 ng of each entry vector containing the promoter, Cas9, terminator, gRNA1, and BASTA resistance were mixed with 250 ng each of the gRNA2 and gRNA3 PCR products, 0.2 µL BSA protein (New England Biolabs, B9000S), 2 µL ligase buffer, 1.2 µL T4 DNA ligase (ThermoFisher, #EL0011), 1 µL BsaI (NEB, #R0535S) and distilled water (to 20 µL). .. To eliminate not fully-ligated intermediate products, we added 0.85 µL ATP (25 mM) and 1 µL Plasmid-Safe ATP-dependent DNase (Epicentre, E3101K) and incubated for 60 min at 37 °C and for 30 min at 70 °C.

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: .. The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA). .. Ten positive clones were picked for DNA sequencing analysis.

    Article Title: Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida
    Article Snippet: .. An aliquot of 7 μl of digested PCR product is incubated with 1 μl of T4 polynucleotide kinase (cat. # EL0014; Thermo Fisher Scientific), 1 μl of T4 DNA ligase (cat. # EK0031; Thermo Fisher Scientific) and 1 μl of ligation buffer for 2 h at room temperature. ..

    DNA Sequencing:

    Article Title: Ablation of myeloid adenosine kinase epigenetically suppresses atherosclerosis in apolipoprotein E-deficient mice
    Article Snippet: The amplified PCR products were cloned into a linearized vector using T4 DNA Ligase (EL0014, Invitrogen, Carlsbad, CA, USA). .. Ten positive clones were picked for DNA sequencing analysis.

    CRISPR:

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: Paragraph title: CRISPR/Cas9‐mediated deletion of enhancer elements and validations ... The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014).

    Sequencing:

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014). .. Successful cloning was validated by Sanger sequencing using a sequencing primer covering the RNU6 promoter region 235 bp 5′ of the Bbs I site.

    Transformation Assay:

    Article Title: Multiplex mutagenesis of four clustered CrRLK1L with CRISPR/Cas9 exposes their growth regulatory roles in response to metal ions
    Article Snippet: For the Golden Gate reaction, 150 ng of the destination vector (pGGZ003) and 250 ng of each entry vector containing the promoter, Cas9, terminator, gRNA1, and BASTA resistance were mixed with 250 ng each of the gRNA2 and gRNA3 PCR products, 0.2 µL BSA protein (New England Biolabs, B9000S), 2 µL ligase buffer, 1.2 µL T4 DNA ligase (ThermoFisher, #EL0011), 1 µL BsaI (NEB, #R0535S) and distilled water (to 20 µL). .. Half of the reaction was transformed into chemically competent E . coli cells (NEB10beta strain) and selected on LB plates containing 100 µg/mL spectinomycin.

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014). .. Afterwards, the digested ligation reaction was transformed into chemically competent E. coli bacteria.

    Synthesized:

    Article Title: Distinct IL‐1α‐responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner
    Article Snippet: For the generation of each enhancer deletion site, a flanking pair of sgRNA constructs was synthesized as DNA oligonucleotides (Eurofins, HPSF, no modifications). .. The double‐stranded and phosphorylated oligos were then diluted 1:8 and ligated into the pX459 vector using the restriction enzyme Bbs I (Bpil, 10 U/μl; Thermo Fisher Scientific, #FD1014) and the T4 DNA ligase (5 U/μl; Thermo Fisher Scientific, #EL0014).

    Hood:

    Article Title: Genome-wide mapping of nucleotide excision repair with XR-seq
    Article Snippet: Handle it in a fume hood and wear protective clothing, gloves and safety goggles. .. T4 DNA ligase (5 U/μl; Thermo Fisher, cat no. 15224041) PEG 8000 (50% (wt/vol), from T4 RNA ligase reaction buffer; NEB, cat. no. B0216L) Anti-(6–4)PP (Cosmo Bio, cat. no. CAC-NM-DND-002) CRITICAL Antibodies from other suppliers may not be as efficient.

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  • 99
    Thermo Fisher t4 dna ligase
    Enzymatic ligation of T. cruzi cap‐4 spliced leader RNA using <t>T4</t> DNA ligase. A) RNA sequences and sequence of the 20‐nt DNA splint; B) HPLC analysis of a typical ligation reaction after 3 h reaction time; reaction conditions: 10 μ m RNA 10 , 12.5 μ m RNA 11 , 12.5 μ m splint; 0.5 m m ATP, 40 m m Tris ⋅ HCl (pH 7.8), 10 m m MgCl 2 , 10 m m DTT, 5 % ( w / v ) PEG 4000, 0.5 U μL −1 T4 DNA ligase; C) LC–ESI mass spectrum of the purified 39‐nt cap‐4 RNA ligation product.
    T4 Dna Ligase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 382 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase/product/Thermo Fisher
    Average 99 stars, based on 382 article reviews
    Price from $9.99 to $1999.99
    t4 dna ligase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher t4 ligase
    Summary of the Golden GATEway cloning kit. A) Outline of the entire cloning procedure. Eight different entry vectors (pGG-EVx) contain different inserts (colored bars). These inserts are assembled in a predefined order using a Golden Gate reaction into Gateway TM  entry vectors at any position (Threeway Gateway TM  cloning). These can then be assembled to establish a final expression vector in an LR reaction. Compatible overhangs are indicated on each Golden Gate entry vector. B) The principle of Golden Gate cloning is illustrated in the top scheme; the principle of Gateway cloning is illustrated in the bottom scheme. Golden Gate cloning utilizes type II restriction endonucleases to generate compatible overhangs that can be ligated with T4 ligase. The ligation of the two compatible inserts from the entry vectors one and two is illustrated. Gateway cloning relies on recombination of specific att sites using a commercially available enzyme mix (LR Clonase II, Life Technologies).
    T4 Ligase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 182 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 ligase/product/Thermo Fisher
    Average 99 stars, based on 182 article reviews
    Price from $9.99 to $1999.99
    t4 ligase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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    Enzymatic ligation of T. cruzi cap‐4 spliced leader RNA using T4 DNA ligase. A) RNA sequences and sequence of the 20‐nt DNA splint; B) HPLC analysis of a typical ligation reaction after 3 h reaction time; reaction conditions: 10 μ m RNA 10 , 12.5 μ m RNA 11 , 12.5 μ m splint; 0.5 m m ATP, 40 m m Tris ⋅ HCl (pH 7.8), 10 m m MgCl 2 , 10 m m DTT, 5 % ( w / v ) PEG 4000, 0.5 U μL −1 T4 DNA ligase; C) LC–ESI mass spectrum of the purified 39‐nt cap‐4 RNA ligation product.

    Journal: Chembiochem

    Article Title: Practical Synthesis of Cap‐4 RNA

    doi: 10.1002/cbic.201900590

    Figure Lengend Snippet: Enzymatic ligation of T. cruzi cap‐4 spliced leader RNA using T4 DNA ligase. A) RNA sequences and sequence of the 20‐nt DNA splint; B) HPLC analysis of a typical ligation reaction after 3 h reaction time; reaction conditions: 10 μ m RNA 10 , 12.5 μ m RNA 11 , 12.5 μ m splint; 0.5 m m ATP, 40 m m Tris ⋅ HCl (pH 7.8), 10 m m MgCl 2 , 10 m m DTT, 5 % ( w / v ) PEG 4000, 0.5 U μL −1 T4 DNA ligase; C) LC–ESI mass spectrum of the purified 39‐nt cap‐4 RNA ligation product.

    Article Snippet: T4 DNA ligase (80 μL; Thermo Fisher, 5 U μL−1 ) was added to a final concentration of 0.5 U μL−1 in a total volume of 0.8 mL before the mixture was incubated for 2 h at 37 °C.

    Techniques: Ligation, Sequencing, High Performance Liquid Chromatography, Purification

    Summary of the Golden GATEway cloning kit. A) Outline of the entire cloning procedure. Eight different entry vectors (pGG-EVx) contain different inserts (colored bars). These inserts are assembled in a predefined order using a Golden Gate reaction into Gateway TM  entry vectors at any position (Threeway Gateway TM  cloning). These can then be assembled to establish a final expression vector in an LR reaction. Compatible overhangs are indicated on each Golden Gate entry vector. B) The principle of Golden Gate cloning is illustrated in the top scheme; the principle of Gateway cloning is illustrated in the bottom scheme. Golden Gate cloning utilizes type II restriction endonucleases to generate compatible overhangs that can be ligated with T4 ligase. The ligation of the two compatible inserts from the entry vectors one and two is illustrated. Gateway cloning relies on recombination of specific att sites using a commercially available enzyme mix (LR Clonase II, Life Technologies).

    Journal: PLoS ONE

    Article Title: Golden GATEway Cloning - A Combinatorial Approach to Generate Fusion and Recombination Constructs

    doi: 10.1371/journal.pone.0076117

    Figure Lengend Snippet: Summary of the Golden GATEway cloning kit. A) Outline of the entire cloning procedure. Eight different entry vectors (pGG-EVx) contain different inserts (colored bars). These inserts are assembled in a predefined order using a Golden Gate reaction into Gateway TM entry vectors at any position (Threeway Gateway TM cloning). These can then be assembled to establish a final expression vector in an LR reaction. Compatible overhangs are indicated on each Golden Gate entry vector. B) The principle of Golden Gate cloning is illustrated in the top scheme; the principle of Gateway cloning is illustrated in the bottom scheme. Golden Gate cloning utilizes type II restriction endonucleases to generate compatible overhangs that can be ligated with T4 ligase. The ligation of the two compatible inserts from the entry vectors one and two is illustrated. Gateway cloning relies on recombination of specific att sites using a commercially available enzyme mix (LR Clonase II, Life Technologies).

    Article Snippet: The 10nM annealed double-stranded oligo dilution was used as an insert in a standard ligation reaction with T4 ligase (5U, Thermo, Fisher)

    Techniques: Clone Assay, Expressing, Plasmid Preparation, Ligation