t4 ligase  (New England Biolabs)


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    Name:
    T4 DNA Ligase
    Description:
    T4 DNA Ligase 100 000 units
    Catalog Number:
    M0202L
    Price:
    256
    Category:
    DNA Ligases
    Size:
    100 000 units
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    Structured Review

    New England Biolabs t4 ligase
    T4 DNA Ligase
    T4 DNA Ligase 100 000 units
    https://www.bioz.com/result/t4 ligase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t4 ligase - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "An Allosteric-Probe for Detection of Alkaline Phosphatase Activity and Its Application in Immunoassay"

    Article Title: An Allosteric-Probe for Detection of Alkaline Phosphatase Activity and Its Application in Immunoassay

    Journal: Frontiers in Chemistry

    doi: 10.3389/fchem.2018.00618

    Response of the AP to ALP, thrombin, T4 ligase, lysozyme, HSA, proteinase K, trypsin, and de-activated ALP for specificity study. The concentration of ALP is 2 U/mL, and the concentration of other enzymes and proteins are 20 U/mL or 10 nM. Error bars indicate the standard deviations of three samples.
    Figure Legend Snippet: Response of the AP to ALP, thrombin, T4 ligase, lysozyme, HSA, proteinase K, trypsin, and de-activated ALP for specificity study. The concentration of ALP is 2 U/mL, and the concentration of other enzymes and proteins are 20 U/mL or 10 nM. Error bars indicate the standard deviations of three samples.

    Techniques Used: ALP Assay, Concentration Assay

    Related Articles

    Plasmid Preparation:

    Article Title:
    Article Snippet: This result is suggestive of substrate inhibition, quite likely due to the nonspecific interaction of the ligase with the DNA. .. Indeed, reaction of 100 nm substrate 1 with 0.1 nm T4 DNA ligase in the presence of added carrier DNA (100 μg/ml pUC19 vector, preincubated 10 min with ligase, ∼150 μm in base pairs) gave a k obs value that was half that found when reacting 100 nm substrate 1 in the absence of added carrier (5 μm in base pairs), giving confidence that the small decrease in turnover rate seen at 1 μm substrate (50 μm base pairs) can be explained primarily by nonspecific inhibition by binding to dsDNA. ..

    Inhibition:

    Article Title:
    Article Snippet: This result is suggestive of substrate inhibition, quite likely due to the nonspecific interaction of the ligase with the DNA. .. Indeed, reaction of 100 nm substrate 1 with 0.1 nm T4 DNA ligase in the presence of added carrier DNA (100 μg/ml pUC19 vector, preincubated 10 min with ligase, ∼150 μm in base pairs) gave a k obs value that was half that found when reacting 100 nm substrate 1 in the absence of added carrier (5 μm in base pairs), giving confidence that the small decrease in turnover rate seen at 1 μm substrate (50 μm base pairs) can be explained primarily by nonspecific inhibition by binding to dsDNA. ..

    Binding Assay:

    Article Title:
    Article Snippet: This result is suggestive of substrate inhibition, quite likely due to the nonspecific interaction of the ligase with the DNA. .. Indeed, reaction of 100 nm substrate 1 with 0.1 nm T4 DNA ligase in the presence of added carrier DNA (100 μg/ml pUC19 vector, preincubated 10 min with ligase, ∼150 μm in base pairs) gave a k obs value that was half that found when reacting 100 nm substrate 1 in the absence of added carrier (5 μm in base pairs), giving confidence that the small decrease in turnover rate seen at 1 μm substrate (50 μm base pairs) can be explained primarily by nonspecific inhibition by binding to dsDNA. ..

    other:

    Article Title: Efficient assembly of very short oligonucleotides using T4 DNA Ligase
    Article Snippet: Specifically, T4 DNA Ligase has been known for some time to be capable of joining oligos as small as pentamers and hexamers on a complete template [ ], however ligases such as Tth DNA Ligase are severely inefficient at the hexamer level [ ].

    Article Title:
    Article Snippet: Given past reports of a weak affinity for non-nicked DNA by T4 DNA ligase, this result may seem surprising.

    Marker:

    Article Title: Efficient strategy for introducing large and multiple changes in plasmid DNA
    Article Snippet: .. Phusion® high-fidelity DNA polymerase, DNA marker, Taq DNA polymerase, T4-PNK, and T4 DNA ligase were purchased from New England Biolabs (Ipswich, MA, USA). .. Human cDNA library was purchased from Clontech Laboratories (Mountain View, CA, USA).

    Ligation:

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases
    Article Snippet: Reactions included 1 μM of the DNA ligase, 100 nM of the substrate and reaction conditions consisting of NEBNext® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)). .. Ligation assays were performed with T3 DNA ligase, T4 DNA ligase, T7 DNA ligase, PBCV1 DNA ligase, hLig3, and E . coli DNA ligase A. .. Experiments were performed in triplicate; the plotted value is the average and the error bars represent the standard deviation across a minimum of 3 replicates. (TIF).

    Article Title: Mitochondrial DNA ligase function in Saccharomyces cerevisiae
    Article Snippet: .. As Figure reveals, while T4 DNA ligase efficiently multimerized dT16 (arrows indicate ligation products in Fig. ), no product was formed following incubation of this substrate with mitochondrial extracts from any of the strains. .. In contrast, Dnl4p ligase activity was detected in a nuclear protein extract prepared from the wild-type yeast strain FF18734 (data not shown).

    Incubation:

    Article Title: A two-nuclease pathway involving RNase H1 is required for primer removal at human mitochondrial OriL
    Article Snippet: .. The reactions were incubated at 37°C for 30 min, after which 1 U of T4 DNA ligase or 300 fmol DNA ligase III was added. .. The reactions were then incubated at 16°C for 18 h and then stopped by addition of 5 μl stop buffer (10 mM Tris–HCl pH 8.0, 0.2 M NaCl, 1 mM EDTA, 660 μg/ml glycogen [Roche] and 100 μg/ml proteinase K [Ambion]) followed by incubation at 42°C for 45 min.

    Article Title: Mitochondrial DNA ligase function in Saccharomyces cerevisiae
    Article Snippet: .. As Figure reveals, while T4 DNA ligase efficiently multimerized dT16 (arrows indicate ligation products in Fig. ), no product was formed following incubation of this substrate with mitochondrial extracts from any of the strains. .. In contrast, Dnl4p ligase activity was detected in a nuclear protein extract prepared from the wild-type yeast strain FF18734 (data not shown).

    Clone Assay:

    Article Title: Improved Method for Rapid and Efficient Determination of Genome Replication and Protein Expression of Clinical Hepatitis B Virus Isolates ▿
    Article Snippet: Besides the two clones described in , seven additional clones generated by the High Fidelityplus DNA polymerase were tested by a transfection experiment as well. .. All nine clones were digested with BspQI and circularized by T4 DNA ligase prior to transfection. ..

    Transfection:

    Article Title: Improved Method for Rapid and Efficient Determination of Genome Replication and Protein Expression of Clinical Hepatitis B Virus Isolates ▿
    Article Snippet: Besides the two clones described in , seven additional clones generated by the High Fidelityplus DNA polymerase were tested by a transfection experiment as well. .. All nine clones were digested with BspQI and circularized by T4 DNA ligase prior to transfection. ..

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    New England Biolabs t4 dna ligase
    Mutations in larger plasmids. ( A ) Schematic representation of LFEAP mutagenesis in large plasmids. The first-round PCRs cut large plasmids into small pieces (~5 kb) with mutagenic ends. The second-round PCRs and the subsequent annealing yield multi-part DNAs with sticky ends, which can be seamlessly joined by <t>T4</t> DNA ligase simultaneously. ( B ) Introduction of mutations in a 25 kb plasmid. Electrophoresis on a 1% agarose gel shows the DNA products generated by the procedure described in the Supplementary Methods. Lanes 1–5: DNA fragments 1 to 5 generated by first-round PCRs. Lane 6: mixture of annealed multi-part DNAs with sticky ends generated by second-round PCRs and the subsequent annealing. Lane 7: the mixture as shown in lane 6 treated with T4 DNA ligase. Lane 8: 1 kb DNA ladder. ( C ) Introduction of mutations in a 50 kb plasmid. Electrophoresis on a 1% agarose gel shows the DNA products generated by the procedure shown in the Supplementary Methods. Lanes 1–10: DNA fragments 1 to 10 generated by first-round PCRs. Lane 11: mixture of annealed multi-part DNAs with sticky ends generated by second-round PCRs and the subsequent annealing. Lane 12: the mixture as shown in lane 11 treated with T4 DNA ligase. Lane 13: 1 kb DNA ladder. ( D ) Electrophoresis on a 0.5% agarose gel of a 25 kb plasmid. Lane 1: 25 kb plasmid before introducing mutations. Lanes 2–6: 25 kb plasmids after introducing mutations propagated from five single colonies. Lane 7: GeneRuler high range DNA ladder (Thermo Fisher Scientific). Incorrect patterns are marked with a ‘×’. ( E .
    T4 Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase/product/New England Biolabs
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    Mutations in larger plasmids. ( A ) Schematic representation of LFEAP mutagenesis in large plasmids. The first-round PCRs cut large plasmids into small pieces (~5 kb) with mutagenic ends. The second-round PCRs and the subsequent annealing yield multi-part DNAs with sticky ends, which can be seamlessly joined by T4 DNA ligase simultaneously. ( B ) Introduction of mutations in a 25 kb plasmid. Electrophoresis on a 1% agarose gel shows the DNA products generated by the procedure described in the Supplementary Methods. Lanes 1–5: DNA fragments 1 to 5 generated by first-round PCRs. Lane 6: mixture of annealed multi-part DNAs with sticky ends generated by second-round PCRs and the subsequent annealing. Lane 7: the mixture as shown in lane 6 treated with T4 DNA ligase. Lane 8: 1 kb DNA ladder. ( C ) Introduction of mutations in a 50 kb plasmid. Electrophoresis on a 1% agarose gel shows the DNA products generated by the procedure shown in the Supplementary Methods. Lanes 1–10: DNA fragments 1 to 10 generated by first-round PCRs. Lane 11: mixture of annealed multi-part DNAs with sticky ends generated by second-round PCRs and the subsequent annealing. Lane 12: the mixture as shown in lane 11 treated with T4 DNA ligase. Lane 13: 1 kb DNA ladder. ( D ) Electrophoresis on a 0.5% agarose gel of a 25 kb plasmid. Lane 1: 25 kb plasmid before introducing mutations. Lanes 2–6: 25 kb plasmids after introducing mutations propagated from five single colonies. Lane 7: GeneRuler high range DNA ladder (Thermo Fisher Scientific). Incorrect patterns are marked with a ‘×’. ( E .

    Journal: Scientific Reports

    Article Title: Efficient strategy for introducing large and multiple changes in plasmid DNA

    doi: 10.1038/s41598-018-20169-8

    Figure Lengend Snippet: Mutations in larger plasmids. ( A ) Schematic representation of LFEAP mutagenesis in large plasmids. The first-round PCRs cut large plasmids into small pieces (~5 kb) with mutagenic ends. The second-round PCRs and the subsequent annealing yield multi-part DNAs with sticky ends, which can be seamlessly joined by T4 DNA ligase simultaneously. ( B ) Introduction of mutations in a 25 kb plasmid. Electrophoresis on a 1% agarose gel shows the DNA products generated by the procedure described in the Supplementary Methods. Lanes 1–5: DNA fragments 1 to 5 generated by first-round PCRs. Lane 6: mixture of annealed multi-part DNAs with sticky ends generated by second-round PCRs and the subsequent annealing. Lane 7: the mixture as shown in lane 6 treated with T4 DNA ligase. Lane 8: 1 kb DNA ladder. ( C ) Introduction of mutations in a 50 kb plasmid. Electrophoresis on a 1% agarose gel shows the DNA products generated by the procedure shown in the Supplementary Methods. Lanes 1–10: DNA fragments 1 to 10 generated by first-round PCRs. Lane 11: mixture of annealed multi-part DNAs with sticky ends generated by second-round PCRs and the subsequent annealing. Lane 12: the mixture as shown in lane 11 treated with T4 DNA ligase. Lane 13: 1 kb DNA ladder. ( D ) Electrophoresis on a 0.5% agarose gel of a 25 kb plasmid. Lane 1: 25 kb plasmid before introducing mutations. Lanes 2–6: 25 kb plasmids after introducing mutations propagated from five single colonies. Lane 7: GeneRuler high range DNA ladder (Thermo Fisher Scientific). Incorrect patterns are marked with a ‘×’. ( E .

    Article Snippet: Phusion® high-fidelity DNA polymerase, DNA marker, Taq DNA polymerase, T4-PNK, and T4 DNA ligase were purchased from New England Biolabs (Ipswich, MA, USA).

    Techniques: Mutagenesis, Plasmid Preparation, Electrophoresis, Agarose Gel Electrophoresis, Generated

    Replication capacity of the HBV genome linearized by ApaI and SphI restriction enzymes. The EcoRI dimer of clone 4B was digested with ApaI or SphI, with or without further treatment with T4 DNA ligase before transfection into Huh7 cells. The uncut dimer

    Journal: Journal of Clinical Microbiology

    Article Title: Improved Method for Rapid and Efficient Determination of Genome Replication and Protein Expression of Clinical Hepatitis B Virus Isolates ▿

    doi: 10.1128/JCM.02340-10

    Figure Lengend Snippet: Replication capacity of the HBV genome linearized by ApaI and SphI restriction enzymes. The EcoRI dimer of clone 4B was digested with ApaI or SphI, with or without further treatment with T4 DNA ligase before transfection into Huh7 cells. The uncut dimer

    Article Snippet: All nine clones were digested with BspQI and circularized by T4 DNA ligase prior to transfection.

    Techniques: Transfection

    Functional characterization of two High Fidelity plus PCR clones of the 4B genome. The two clones were transfected directly (lanes 1 and 4) following digestion with BspQI (lanes 2 and 5) or BspQI digestion plus treatment with T4 DNA ligase (lanes 3 and

    Journal: Journal of Clinical Microbiology

    Article Title: Improved Method for Rapid and Efficient Determination of Genome Replication and Protein Expression of Clinical Hepatitis B Virus Isolates ▿

    doi: 10.1128/JCM.02340-10

    Figure Lengend Snippet: Functional characterization of two High Fidelity plus PCR clones of the 4B genome. The two clones were transfected directly (lanes 1 and 4) following digestion with BspQI (lanes 2 and 5) or BspQI digestion plus treatment with T4 DNA ligase (lanes 3 and

    Article Snippet: All nine clones were digested with BspQI and circularized by T4 DNA ligase prior to transfection.

    Techniques: Functional Assay, Polymerase Chain Reaction, Clone Assay, Transfection

    Yeast mitochondrial protein extracts ligate nicked DNA:DNA substrates. Mitochondrial protein extracts were prepared from strain 2780-49BWT, cdc9-1 ts and Δ dnl4 yeast and their ability to seal a nick in a DNA:DNA duplex substrate was assayed. Reactions were carried out as described in Materials and Methods, with extracts preincubated at the temperatures indicated. The lower band indicates unligated substrate DNA, while the upper band (arrow) indicates the ligated DNA product. The positive control (+) is substrate plus T4 DNA ligase; the negative control (–) is substrate alone.

    Journal: Nucleic Acids Research

    Article Title: Mitochondrial DNA ligase function in Saccharomyces cerevisiae

    doi:

    Figure Lengend Snippet: Yeast mitochondrial protein extracts ligate nicked DNA:DNA substrates. Mitochondrial protein extracts were prepared from strain 2780-49BWT, cdc9-1 ts and Δ dnl4 yeast and their ability to seal a nick in a DNA:DNA duplex substrate was assayed. Reactions were carried out as described in Materials and Methods, with extracts preincubated at the temperatures indicated. The lower band indicates unligated substrate DNA, while the upper band (arrow) indicates the ligated DNA product. The positive control (+) is substrate plus T4 DNA ligase; the negative control (–) is substrate alone.

    Article Snippet: As Figure reveals, while T4 DNA ligase efficiently multimerized dT16 (arrows indicate ligation products in Fig. ), no product was formed following incubation of this substrate with mitochondrial extracts from any of the strains.

    Techniques: Positive Control, Negative Control

    Yeast mitochondrial protein extracts do not ligate nicked DNA:RNA substrates. DNA ligase reactions were carried out on mitochondrial protein extracts in a manner similar to Figure 1, however, the substrate was a DNA:RNA duplex. The lower band (dT 16 ) indicates unligated substrate, while the upper bands (dT 32 , dT 48 and dT 64 ) indicate products. The positive control (+) is substrate plus T4 DNA ligase; the negative control (–) is substrate alone.

    Journal: Nucleic Acids Research

    Article Title: Mitochondrial DNA ligase function in Saccharomyces cerevisiae

    doi:

    Figure Lengend Snippet: Yeast mitochondrial protein extracts do not ligate nicked DNA:RNA substrates. DNA ligase reactions were carried out on mitochondrial protein extracts in a manner similar to Figure 1, however, the substrate was a DNA:RNA duplex. The lower band (dT 16 ) indicates unligated substrate, while the upper bands (dT 32 , dT 48 and dT 64 ) indicate products. The positive control (+) is substrate plus T4 DNA ligase; the negative control (–) is substrate alone.

    Article Snippet: As Figure reveals, while T4 DNA ligase efficiently multimerized dT16 (arrows indicate ligation products in Fig. ), no product was formed following incubation of this substrate with mitochondrial extracts from any of the strains.

    Techniques: Positive Control, Negative Control

    Schematic representation of DNA ligase fusions. All DNA ligases contain a catalytic core NTase domain (blue) and an OBD (red), which are fairly well conserved. Many ligases also have additional domains, such as the N-terminal ZnF (yellow) and DBD (green) in Human Lig3 and the N-terminal domain (NTD) of T4 DNA ligase (purple). Wild type PBCV1 ligase, which contains only the core NTase and OBD domains, was chosen for fusion to other binding domains: Sso7d (white) at both the N- and C-termini, the hLig3 ZnF domain, and the T4 DNA ligase NTD.

    Journal: PLoS ONE

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases

    doi: 10.1371/journal.pone.0190062

    Figure Lengend Snippet: Schematic representation of DNA ligase fusions. All DNA ligases contain a catalytic core NTase domain (blue) and an OBD (red), which are fairly well conserved. Many ligases also have additional domains, such as the N-terminal ZnF (yellow) and DBD (green) in Human Lig3 and the N-terminal domain (NTD) of T4 DNA ligase (purple). Wild type PBCV1 ligase, which contains only the core NTase and OBD domains, was chosen for fusion to other binding domains: Sso7d (white) at both the N- and C-termini, the hLig3 ZnF domain, and the T4 DNA ligase NTD.

    Article Snippet: Ligation assays were performed with T3 DNA ligase, T4 DNA ligase, T7 DNA ligase, PBCV1 DNA ligase, hLig3, and E . coli DNA ligase A.

    Techniques: Binding Assay

    Wild type DNA ligase λ DNA digest ligation assay. Agarose gel electrophoresis of λ DNA cut by EcoRV (A/T Blunt, 1 ), NruI (G/C Blunt, 2 ), BstNI (5′ SBO, 3 ), Hpy188I (3′SBO, 4 ), NdeI (2 BO, 5 ) and BamHI (4 BO, 6 ), generating DNA fragments with ligatable ends. 0.5 ng of the cut DNA was ligated in the presence of T4 ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl2, 1 mM DTT, 1 mM ATP, 6% polyethylene glycol (PEG 6000)) and 7 μM of the indicated DNA ligase for 1 hour at 25°C. Ligation assays performed with T4 DNA ligase (A), T3 DNA ligase (B), PBCV1 DNA ligase (C) and, hLig3 (D), respectively. E) Gel of restriction enzyme digested λ DNA samples as well as a schematic depiction of each substrate. The DNA fragments were visualized using ethidium bromide stain.

    Journal: PLoS ONE

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases

    doi: 10.1371/journal.pone.0190062

    Figure Lengend Snippet: Wild type DNA ligase λ DNA digest ligation assay. Agarose gel electrophoresis of λ DNA cut by EcoRV (A/T Blunt, 1 ), NruI (G/C Blunt, 2 ), BstNI (5′ SBO, 3 ), Hpy188I (3′SBO, 4 ), NdeI (2 BO, 5 ) and BamHI (4 BO, 6 ), generating DNA fragments with ligatable ends. 0.5 ng of the cut DNA was ligated in the presence of T4 ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl2, 1 mM DTT, 1 mM ATP, 6% polyethylene glycol (PEG 6000)) and 7 μM of the indicated DNA ligase for 1 hour at 25°C. Ligation assays performed with T4 DNA ligase (A), T3 DNA ligase (B), PBCV1 DNA ligase (C) and, hLig3 (D), respectively. E) Gel of restriction enzyme digested λ DNA samples as well as a schematic depiction of each substrate. The DNA fragments were visualized using ethidium bromide stain.

    Article Snippet: Ligation assays were performed with T3 DNA ligase, T4 DNA ligase, T7 DNA ligase, PBCV1 DNA ligase, hLig3, and E . coli DNA ligase A.

    Techniques: Ligation, Agarose Gel Electrophoresis, Staining

    Wild type DNA ligase blunt/cohesive capillary electrophoresis assay. Bar graphs depict the fraction of either ligated DNA (product, blue) or abortive adenylylation (App, red) produced in a 20-minute sealing reaction with the indicated DNA substrate. Reactions included 1 μM of the DNA ligase, 100 nM of the substrate and reaction conditions consisting of either T4 DNA ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl 2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)). Ligation assays performed with T4 DNA ligase (A), T3 DNA ligase (B), PBCV1 DNA ligase (C) and hLig3 (D), respectively Experiments were performed in triplicate; the plotted value is the average and the error bars represent the standard deviation across replicates.

    Journal: PLoS ONE

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases

    doi: 10.1371/journal.pone.0190062

    Figure Lengend Snippet: Wild type DNA ligase blunt/cohesive capillary electrophoresis assay. Bar graphs depict the fraction of either ligated DNA (product, blue) or abortive adenylylation (App, red) produced in a 20-minute sealing reaction with the indicated DNA substrate. Reactions included 1 μM of the DNA ligase, 100 nM of the substrate and reaction conditions consisting of either T4 DNA ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl 2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)). Ligation assays performed with T4 DNA ligase (A), T3 DNA ligase (B), PBCV1 DNA ligase (C) and hLig3 (D), respectively Experiments were performed in triplicate; the plotted value is the average and the error bars represent the standard deviation across replicates.

    Article Snippet: Ligation assays were performed with T3 DNA ligase, T4 DNA ligase, T7 DNA ligase, PBCV1 DNA ligase, hLig3, and E . coli DNA ligase A.

    Techniques: Electrophoresis, Produced, Ligation, Standard Deviation

    Effect of DBD on blunt/cohesive end ligation. Bar graphs depict the fraction of either ligated DNA (product, blue) or abortive adenylylation (App, red) produced in a 20-minute sealing reaction with the indicated DNA substrate. Reactions included 1 μM of the DNA ligase, 100 nM of the substrate and reaction conditions consisting of either T4 DNA ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl 2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)). Ligation assays performed with PBCV1-Nterm-Sso7d (A), PBCV1-Cterm-Sso7d terminus (B), PBCV1-Nterm-ZnF (C), PBCV1-Nterm-T4NTD (D). Experiments were performed in triplicate; the plotted value is the average and the error bars represent the standard deviation across replicates.

    Journal: PLoS ONE

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases

    doi: 10.1371/journal.pone.0190062

    Figure Lengend Snippet: Effect of DBD on blunt/cohesive end ligation. Bar graphs depict the fraction of either ligated DNA (product, blue) or abortive adenylylation (App, red) produced in a 20-minute sealing reaction with the indicated DNA substrate. Reactions included 1 μM of the DNA ligase, 100 nM of the substrate and reaction conditions consisting of either T4 DNA ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl 2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)). Ligation assays performed with PBCV1-Nterm-Sso7d (A), PBCV1-Cterm-Sso7d terminus (B), PBCV1-Nterm-ZnF (C), PBCV1-Nterm-T4NTD (D). Experiments were performed in triplicate; the plotted value is the average and the error bars represent the standard deviation across replicates.

    Article Snippet: Ligation assays were performed with T3 DNA ligase, T4 DNA ligase, T7 DNA ligase, PBCV1 DNA ligase, hLig3, and E . coli DNA ligase A.

    Techniques: Ligation, Produced, Standard Deviation